==================================BSR46================================== 46. Apolipoproteins crossreferenced with: - genomic sequence, or sequence (alone), or nucleotide sequence, or nucleic acid sequence, or base sequence and Repetitive sequences (alone) - Do not crossreference with apolipo- proteins. 1 TI - Mouse apolipoprotein A-IV gene: nucleotide sequence and induction by a high-lipid diet. 2 TI - On computer-assisted analysis of biological sequences: proline punctuation, consensus sequences, and apolipoprotein repeats. 3 TI - Isolation, expression and characterization of a human apolipoprotein B 100-specific cDNA clone. 4 TI - Conserved sequence motifs upstream from the co-ordinately expressed vitellogenin and apoVLDLII genes of chicken. 5 TI - Cloning and expression of human apolipoprotein D cDNA. prt ar compr include mh 1 UI - 87089722 AU - Williams SC ; Bruckheimer SM ; Lusis AJ ; LeBoeuf RC ; Kinniburgh AJ TI - Mouse apolipoprotein A-IV gene: nucleotide sequence and induction by a high-lipid diet. AB - Apolipoprotein A-IV (apo A-IV) functions in conjunction with other apolipoproteins to form lipoprotein particles which are involved in lipid homeostasis. In this report we present the nucleotide sequence of the mouse apo A-IV gene and demonstrate its induction in the liver by chronically high dietary lipid. The apo A-IV gene consists of three exons and two introns. The introns separate evolutionarily conserved and functional polypeptide domains. Intron 1 divides most of the apo A-IV signal peptide from the amino terminus of the mature plasma protein. The second intron separates a highly evolutionarily conserved, variant amphipathic peptide repeat from the remainder of the mature apo A-IV protein. The 5' flanking region has several interesting features. The apo A-IV gene has variant TATA and CAT box sequences, TTTAAA and CCAACG, respectively. There are five G-rich direct repeats of 10 nucleotides and a short inverted repeat in the 5' flanking region. We speculate that these sequence elements in the 5' flanking region may be involved in the regulation of apo A-IV gene expression. We also show that chronically high dietary lipid induces liver apo A-IV levels 10-fold in C57BL/6 mice, a strain susceptible to atherosclerotic lesions, while we observed no induction in nonsusceptible BALB/c and C3H mice. MH - Amino Acid Sequence ; Animal ; Apolipoproteins A/BIOSYNTHESIS/*GENETICS ; Base Sequence ; Cloning, Molecular ; Comparative Study ; Dietary Fats/ *PHARMACOLOGY ; DNA/METABOLISM ; Exons ; *Genes, Structural/DRUG EFFECTS ; Human ; Mice ; Promoter Regions (Genetics) ; Rats ; Sequence Homology, Nucleic Acid ; Species Specificity ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - Mol Cell Biol 1986 Nov;6(11):3807-14 2 UI - 87085187 AU - Boguski MS ; Freeman M ; Elshourbagy NA ; Taylor JM ; Gordon JI TI - On computer-assisted analysis of biological sequences: proline punctuation, consensus sequences, and apolipoprotein repeats. AB - During the past several years, the use of computer programs in the analysis of protein and DNA sequences has become commonplace. In all but the simplest procedures, the ability to critically review the results obtained with computer methods requires a basic knowledge of the algorithms employed (and the assumptions upon which they are based), an awareness of the capabilities and limitations of the particular program that implements an algorithm, and some familiarity with probability and statistics. We describe a number of computer methods that have been applied to the analysis of apolipoprotein sequences. We discuss the suitability of these methods for particular problems, how the choice of initial "parameters: can affect the results, and what the results can tell us about protein or gene sequences. We also identify some outstanding problems of apolipoprotein sequence analysis where further work is needed. RF - REVIEW ARTICLE: 97 REFS. MH - *Amino Acid Sequence ; *Apolipoproteins ; Base Sequence ; DNA ; Human ; Information Systems ; Phylogeny ; Proline ; Repetitive Sequences, Nucleic Acid ; Review ; *Software ; Structure-Activity Relationship ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - J Lipid Res 1986 Oct;27(10):1011-34 3 UI - 87076044 AU - Pfitzner R ; Wagener R ; Stoffel W TI - Isolation, expression and characterization of a human apolipoprotein B 100-specific cDNA clone. AB - The isolation and characterization of a human apolipoprotein B 100-specific cDNA clone (lambda gt-B1) containing a 1321 base pairs (bp) spanning insert is described. It encodes the 3'-nontranslated 281 bp long region up to the polyadenylation site and 1040 bp of the C-terminal coding region of 345 amino-acid residues of human apo B 100 and the stop codon. The lambda gt-B1 cDNA clone has been isolated from a human hepatoma cDNA expression library by immunoscreening using affinity-purified polyclonal anti apo B 100 antibodies. The nucleotide sequence of the apo B 100 insert has been determined. A part of the polypeptide sequence derived from this nucleotide sequence was identical with the amino-acid sequence obtained by protein sequencing of a purified cyanogen bromide fragment of apo B 100. The fusion protein consisting of beta-galactosidase and the 345 amino-acid residue long C-terminus of apo B 100 had an apparent molecular mass of 148 kDa in NaDodSO4 polyacrylamide gel electrophoresis. In Northern blot hybridization analysis the insert of the apo B 100-cDNA clone hybridized to a 20 to 22 kb mRNA from adult human liver. MH - Amino Acid Sequence ; Apolipoproteins B/*GENETICS ; Base Sequence ; Chromosome Mapping ; Cloning, Molecular ; DNA/*GENETICS/ISOLATION & PURIFICATION ; DNA Restriction Enzymes ; Human ; Support, Non-U.S. Gov't SO - Biol Chem Hoppe Seyler 1986 Oct;367(10):1077-83 4 UI - 87066737 AU - van het Schip F ; Strijker R ; Samallo J ; Gruber M ; Geert AB TI - Conserved sequence motifs upstream from the co-ordinately expressed vitellogenin and apoVLDLII genes of chicken. AB - The vitellogenin and apoVLDLII yolk protein genes of chicken are transcribed in the liver upon estrogenization. To get information on putative regulatory elements, we compared more than 2 kb of their 5' flanking DNA sequences. Common sequence motifs were found in regions exhibiting estrogen-induced changes in chromatin structure. Stretches of alternating pyrimidines and purines of about 30-nucleotides long are present at roughly similar positions. A distinct box of sequence homology in the chicken genes also appears to be present at a similar position in front of the vitellogenin genes of Xenopus laevis, but is absent from the estrogen-responsive egg-white protein genes expressed in the oviduct. In front of the vitellogenin (position -595) and the VLDLII gene (position -548), a DNA element of about 300 base-pairs was found, which possesses structural characteristics of a mobile genetic element and bears homology to the transposon-like Vi element of Xenopus laevis. MH - Animal ; Apolipoproteins/*GENETICS ; Base Sequence ; Chickens ; DNA Restriction Enzymes ; Female ; *Genes, Structural ; Lipoproteins, VLDL/ *GENETICS ; Liver/METABOLISM ; Oviducts/METABOLISM ; Support, Non-U.S. Gov't ; Transcription, Genetic ; *Vitellogenin/*GENETICS ; Xenopus SO - Nucleic Acids Res 1986 Nov 11;14(21):8669-80 5 UI - 87057347 AU - Drayna D ; Fielding C ; McLean J ; Baer B ; Castro G ; Chen E ; Comstock L ; Henzel W ; Kohr W ; Rhee L ; et al TI - Cloning and expression of human apolipoprotein D cDNA. AB - The amino acid sequence of human apolipoprotein D, a component of high density lipoprotein, has been obtained from the cloned cDNA sequence. The 169-amino acid protein has no marked similarity to other apolipoprotein sequences, but has a high degree of homology to plasma retinol-binding protein and other members of the alpha 2u-globulin protein superfamily. Apolipoprotein D mRNA has been detected in human liver, intestine, pancreas, kidney, placenta, adrenal, spleen, and fetal brain tissue. Tissue culture cells transfected with the cloned cDNA secrete material that reacts with anti-apoD antibodies. MH - Amino Acid Sequence ; Apolipoproteins/BLOOD/*GENETICS ; Base Sequence ; *Cloning, Molecular ; DNA/*METABOLISM ; Human ; Molecular Weight ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; *Transcription, Genetic SO - J Biol Chem 1986 Dec 15;261(35):16535-9 6 UI - 87057153 AU - Blackhart BD ; Ludwig EM ; Pierotti VR ; Caiati L ; Onasch MA ; Wallis SC ; Powell L ; Pease R ; Knott TJ ; Chu ML ; et al TI - Structure of the human apolipoprotein B gene. AB - Human apolipoprotein B100 cDNA is 14 kilobases in length and encodes a 4563-amino acid precursor protein. The corresponding human gene has been isolated as a series of overlapping lambda clones and extends over 43 kilobases. The gene comprises 29 exons and 28 introns. The distribution of introns is extremely asymmetrical, most of them appearing in the 5'-terminal one-third of the gene. Although most of the exons fall within the normal size limits for mammalian genes, two are unusually long: 1906 and 7572 base pairs. The latter exon is by far the longest reported for a vertebrate gene. MH - Apolipoproteins B/*GENETICS ; Base Sequence ; DNA/GENETICS ; DNA, Recombinant ; Exons ; Human ; Introns ; Nucleic Acid Hybridization ; Phage Lambda/GENETICS ; Polymorphism (Genetics) ; Repetitive Sequences, Nucleic Acid SO - J Biol Chem 1986 Nov 25;261(33):15364-7 7 UI - 87055262 AU - Strijker R ; Blom van Assendelft G ; Dikkeschei BD ; Gruber M ; Ab G TI - Estradiol-dependent transcription initiation upstream from the chicken apoVLDLII gene coding for the very-low-density apolipoprotein II. AB - We have investigated RNAs originating from the 5'-flanking region of the chicken very-low-density apolipoprotein II (apoVLDLII) gene. S1 nuclease mapping and primer extension experiments revealed two minor upstream transcription start points located 1105 and 1530 nucleotides in front of the apoVLDLII gene. Transcription starting at these points is dependent upon estradiol as is transcription from the major start points. The transcripts are polyadenylated, but are not detectable in polysomes. Run-on assays indicated that the low concentration of the upstream initiated transcripts is due both to low transcription levels and to low transcript stability. The sequence around the upstream start points does not show strong homologies with consensus sequences of promoters for eukaryotic protein encoding genes. Nevertheless, the upstream sequences are transcribed in vivo by RNA polymerase II. MH - Animal ; Apolipoproteins/*GENETICS ; Base Sequence ; Chickens ; Comparative Study ; Estradiol/*PHARMACOLOGY ; Female ; Lipoproteins, VLDL/ *GENETICS ; Male ; Promoter Regions (Genetics) ; RNA Polymerase II/ METABOLISM ; RNA, Messenger/BIOSYNTHESIS/PHYSIOLOGY ; Sequence Homology, Nucleic Acid ; Support, Non-U.S. Gov't ; Transcription, Genetic/*DRUG EFFECTS SO - Gene 1986;45(1):27-35 8 UI - 87049597 AU - Cladaras C ; Hadzopoulou-Cladaras M ; Avila R ; Nussbaum AL ; Nicolosi R ; Zannis VI TI - Complementary DNA derived structure of the amino-terminal domain of human apolipoprotein B and size of its messenger RNA transcript. AB - In this paper the sequence of a 5.2-kilobase (kb) cDNA covering the amino-terminal domain of human apolipoprotein B-100 (apoB-100) is reported. The cDNA-derived protein sequence provides the primary structure of 1748 amino acids. This segment of apoB-100 is more hydrophilic than hydrophobic and contains short stretches of predicted helical and beta structures that are interrupted by beta turns. Blotting analysis of RNA isolated from fetal human and adult monkey tissues and various human cell lines showed synthesis of apoB mRNA by liver and intestine and by cells of hepatic (HepG2) and intestinal (Caco-2) origin. The isolation and characterization of overlapping cDNA clones, which provide a nearly full-length copy of human apoB-100, are also reported. From the length of these clones the size of the cytoplasmic apoB mRNA is estimated to be 14.0 kb and codes for a protein of approximately 512,000 daltons. MH - Adult ; Amino Acid Sequence ; Apolipoproteins B/*GENETICS ; Base Sequence ; DNA/*ANALYSIS ; DNA Restriction Enzymes ; *Genes, Structural ; Human ; Liver/METABOLISM ; RNA, Messenger/*GENETICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; *Transcription, Genetic SO - Biochemistry 1986 Sep 23;25(19):5351-7 9 UI - 87041474 AU - Karathanasis SK ; Oettgen P ; Haddad IA ; Antonarakis SE TI - Structure, evolution, and polymorphisms of the human apolipoprotein A4 gene (APOA4). AB - The genes coding for three proteins of the plasma lipid transport system--apolipoproteins A1 (APOA1), C3 (APOC3), and A4 (APOA4)--are closely linked and tandemly organized on the long arm of human chromosome 11. In this study the human APOA4 gene has been isolated and characterized. In contrast to APOA1 and APOC3 genes, which contain three introns, the APOA4 gene contains only two. An intron interrupting the 5' noncoding region of the APOA1 and APOC3 mRNAs is absent from the corresponding position of the APOA4 mRNA. However, similar to APOA1 and APOC3 genes, the introns of the APOA4 gene separate nucleotide sequences coding for the signal peptide and the amphipathic domains in APOA4. These results suggest that the APOA1, APOC3, and APOA4 genes were derived from a common evolutionary ancestor and indicate that during evolution the APOA4 gene lost one of its ancestral introns. Two restriction endonuclease sites, an Xba I located in the second intron of the APOA4 gene and a different Xba I located 9 kilobases 3' to the APOA4 gene, are polymorphic in Mediterranean and Northern European populations. Haplotype analysis indicated that even though these polymorphic sites are located within 9 kilobases they do not display significant nonrandom association. Finally, restriction mapping analysis of DNA from a patient with combined APOA1-APOC3 deficiency and premature coronary artery disease indicated that this patient has a structurally normal APOA4 gene. MH - Apolipoproteins A/DEFICIENCY/*GENETICS ; Apolipoproteins C/DEFICIENCY/ GENETICS ; Base Sequence ; Coronary Disease/FAMILIAL & GENETIC ; DNA/ ANALYSIS ; Evolution ; Human ; *Polymorphism (Genetics) ; Recombination, Genetic ; RNA, Messenger/ANALYSIS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Nov;83(22):8457-61 10 UI - 87041416 AU - Law SW ; Grant SM ; Higuchi K ; Hospattankar A ; Lackner K ; Lee N ; Brewer HB Jr TI - Human liver apolipoprotein B-100 cDNA: complete nucleic acid and derived amino acid sequence. AB - Human apolipoprotein B-100 (apoB-100), the ligand on low density lipoproteins that interacts with the low density lipoprotein receptor and initiates receptor-mediated endocytosis and low density lipoprotein catabolism, has been cloned, and the complete nucleic acid and derived amino acid sequences have been determined. ApoB-100 cDNAs were isolated from normal human liver cDNA libraries utilizing immunoscreening as well as filter hybridization with radiolabeled apoB-100 oligodeoxynucleotides. The apoB-100 mRNA is 14.1 kilobases long encoding a mature apoB-100 protein of 4536 amino acids with a calculated amino acid molecular weight of 512,723. ApoB-100 contains 20 potential glycosylation sites, and 12 of a total of 25 cysteine residues are located in the amino-terminal region of the apolipoprotein providing a potential globular structure of the amino terminus of the protein. ApoB-100 contains relatively few regions of amphipathic helices, but compared to other human apolipoproteins it is enriched in beta-structure. The delineation of the entire human apoB-100 sequence will now permit a detailed analysis of the conformation of the protein, the low density lipoprotein receptor binding domain(s), and the structural relationship between apoB-100 and apoB-48 and will provide the basis for the study of genetic defects in apoB-100 in patients with dyslipoproteinemias. MH - Amino Acid Sequence ; Apolipoproteins B/ANALYSIS/*GENETICS ; Base Sequence ; DNA/*ANALYSIS/ISOLATION & PURIFICATION ; Human ; Liver/ *ANALYSIS ; Protein Conformation ; RNA, Messenger/ANALYSIS SO - Proc Natl Acad Sci USA 1986 Nov;83(21):8142-6 11 UI - 87039352 AU - Yang CY ; Chen SH ; Gianturco SH ; Bradley WA ; Sparrow JT ; Tanimura M ; Li WH ; Sparrow DA ; DeLoof H ; Rosseneu M ; et al TI - Sequence, structure, receptor-binding domains and internal repeats of human apolipoprotein B-100. AB - Apolipoprotein (apo) B-100, the major protein component in low density lipoprotein (LDL), is the ligand that binds to the LDL receptor. It is important in the metabolism of LDL and elevated plasma levels of LDL-apo B are strongly associated with increased risk of coronary artery disease. Although apo B-100 is of great clinical and biological importance its primary structure has defied chemical elucidation, mainly because of its enormous size, insolubility, and tendency to aggregate. Less than 5% of the apo B-100 sequence has been reported, despite the efforts of many laboratories over the past twenty years. Here we report the complete amino acid sequence of human apo B-100 as deducted by sequence analysis of complementary DNA clones; 2,366 of the 4,536 residues were also confirmed by direct sequencing of apo B-100 tryptic peptides. The distribution of trypsin-accessible and -inaccessible peptides of the protein on LDL is non-random and they can be grouped into 5 hypothetical domains. Of 20 potential N-glycosylation sites identified in the sequence, 13 were found by direct peptide sequencing to be glycosylated, and 4 unglycosylated. Examination of the primary structure of apo B-100 reveals that it contains a large number of long (greater than 70 residues) internal repeats and an even larger number of shorter ones, suggesting that the apo B-100 sequence was derived largely from internal duplications. Finally, using synthetic peptides of a specific region of apo B-100, we have identified a potential LDL receptor-binding domain (residues 3,345-3,381) which can bind to the LDL receptor and suppress 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase activities in cultured human fibroblasts. MH - Amino Acid Sequence ; Apolipoproteins A/METABOLISM ; Apolipoproteins B/ *ANALYSIS/GENETICS ; Human ; Protein Conformation ; Receptors, LDL/ *METABOLISM ; Repetitive Sequences, Nucleic Acid ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Nature 1986 Oct 23-29;323(6090):738-42 12 UI - 87039351 AU - Knott TJ ; Pease RJ ; Powell LM ; Wallis SC ; Rall SC Jr ; Innerarity TL ; Blackhart B ; Taylor WH ; Marcel Y ; Milne R ; et al TI - Complete protein sequence and identification of structural domains of human apolipoprotein B. AB - Epidemiological, pathological and genetic studies show a strong positive correlation between elevated plasma concentrations of low-density lipoprotein (LDL) cholesterol and the risk of premature coronary heart disease. Apolipoprotein (apo) B-100 is the sole protein component of LDL and is the ligand responsible for the receptor-mediated uptake and clearance of LDL from the circulation. Apo B-100 is made by the liver and is essential for the assembly of triglyceride-rich very low-density lipoproteins (VLDL) in the cisternae of the endoplasmic reticulum and for their secretion into the plasma. VLDL transports triglyceride to peripheral muscle and adipose tissue, where the triglyceride is hydrolysed by lipoprotein lipase. The resultant particle, relatively enriched in cholesteryl ester, constitutes LDL. LDL delivers cholesterol to peripheral tissues where it is used for membrane and steroid hormone biosynthesis and to the liver, the only organ which can catabolize and excrete cholesterol. Plasma LDL levels are therefore determined by the balance between their rate of production from VLDL and clearance by the hepatic LDL (apo B/E) receptor pathway. Here we report the complete 4,563-amino-acid sequence of apo B-100 precursor (relative molecular mass (Mr) 514,000 (514K] determined from complementary DNA clones. Numerous lipid-binding structures are distributed throughout the extraordinary length of apo B-100 and must underlie its special functions as a nucleus for lipoprotein assembly and maintenance of plasma lipoprotein integrity. A domain enriched in basic amino-acid residues has been identified as important for the cellular uptake of cholesterol by the LDL receptor pathway. MH - Amino Acid Sequence ; Antibodies, Monoclonal ; Apolipoproteins B/ *ANALYSIS/GENETICS ; Base Sequence ; Human ; Lipoproteins, LDL/METABOLISM ; Receptors, LDL/METABOLISM SO - Nature 1986 Oct 23-29;323(6090):734-8 13 UI - 87033692 AU - Folz RJ ; Gordon JI TI - Deletion of the propeptide from human preproapolipoprotein A-II redirects cotranslational processing by signal peptidase. AB - The functions of NH2-terminal propeptides are not known. We have used apoA-II as a model to study prosegment structure/function relationships. The primary translation product of human apolipoprotein A-II mRNA contains an 18-amino acid signal peptide, a 5-amino acid propeptide, and the mature 77-amino acid plasma protein sequence. Its propeptide was deleted by site-directed mutagenesis of a cloned cDNA. The effects of this mutation on cotranslational translocation and proteolytic processing were assessed using an in vitro transcription/translation/microsomal membrane processing system. Deletion of the propeptide did not affect cotranslational translocation. However, without its propeptide, signal peptidase cleavage was redirected to a different site located between the 2nd and 3rd residues of the mature protein. Since the primary structure of the signal peptide was not altered in the mutant, these results suggest that sequences located downstream from the signal peptidase cleavage site (e.g. in propeptides) may modulate, or participate in defining, the correct site of cotranslational proteolytic processing. MH - Apolipoproteins A/*GENETICS ; Base Sequence ; *Chromosome Deletion ; Cloning, Molecular ; DNA/METABOLISM ; Genes, Structural ; Human ; Liver/ METABOLISM ; Models, Genetic ; Mutation ; Peptide Peptidohydrolases/ *METABOLISM ; Plasmids ; Protein Precursors/*GENETICS ; RNA, Messenger/ *GENETICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Transcription, Genetic ; *Translation, Genetic SO - J Biol Chem 1986 Nov 5;261(31):14752-9 14 UI - 87010278 AU - Nagashima M ; Morris G ; Howlett G ; Fidge N ; Schreiber G TI - Amino acid sequence of rat apolipoprotein A-II deduced from the nucleotide sequence of cloned cDNA. AB - A rat apolipoprotein A-II cDNA clone was isolated from a rat liver cDNA library by in situ hybridization of bacteriophage plaques using a 32P-labeled human apoA-II cDNA as a probe. The cDNA insert from this clone was characterized by DNA sequencing. The amino acid composition derived from the DNA sequence data matched well with that of rat apoA-II reported earlier (Herbert et al. 1974. J. Biol Chem. 249: 5718-5724), indicating that the cDNA insert coded for rat apoA-II. Further evidence was provided by a comparison of the amino acid sequence of rat apoA-II obtained here with that of human apoA-II (Brewer et al. 1972. Proc. Natl. Acad. Sci. USA. 69: 1304-1308). While the rat apoA-II cDNA insert did not code for the entire presegment, it had the same COOH-terminal residues of the presegment as well as the same prosegment (Ala-Leu-Val-Arg-Arg) as in human preproapoA-II, suggesting that rat apoA-II was also synthesized initially as preproapoA-II. Mature rat apoA-II contains 79 amino acids. Residue 6 of mature rat apoA-II is Asp, while it is Cys in human apoA-II, and this would account for the absence of dimeric forms of rat apoA-II in plasma. While the overall amino acid sequence homology between rat and human apoA-II is about 50%, the amphipathic alpha-helical structures, which are responsible for lipid-binding, seem to be conserved in the two proteins. The size of rat apoA-II mRNA was estimated to be about 600 nucleotides.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Amino Acid Sequence ; Animal ; Apolipoproteins A/*GENETICS ; Base Sequence ; *Cloning, Molecular ; Comparative Study ; DNA/*METABOLISM ; Human ; Liver/*METABOLISM ; Molecular Weight ; Nucleic Acid Hybridization ; Rats ; RNA, Messenger/GENETICS ; Sequence Homology, Nucleic Acid ; Species Specificity ; Support, Non-U.S. Gov't SO - J Lipid Res 1986 Jul;27(7):706-12 15 UI - 87008617 AU - Fung WP ; Howlett GJ ; Schreiber G TI - Structure and expression of the rat apolipoprotein E gene. AB - The rat apolipoprotein E gene was isolated from a genomic library by screening with a cDNA probe. The nucleotide sequence of the gene plus a 5' flanking region of 623 nucleotides and a 3' flanking region of 234 nucleotides was determined. The gene contained three introns, which were located in the 5' nontranslated region and in the regions coding for the signal peptide and the mature protein. A "TATA box: sequence TATAATT was found beginning at nucleotide -32. Two copies of the sequences GGGCGG or CCGCCC, potential binding sites for the transcription protein factor Sp 1, were found adjacent to the TATA box, beginning at nucleotides -52 and -72. The 5' proximal flanking region, up to about 140 nucleotides, was found to be highly conserved (82% homology) in the gene for rat and human. This region was GC-rich (68% G + C) and contained self-complementary sequences with the potential to form hairpin loop structures, suggesting a regulatory role of this region in gene transcription. Two open reading frames with promoter sequences were found, one located in the first and the other in the second intron. Expression of rat apolipoprotein E mRNA in murine L cells transfected with the cloned gene was detected at levels comparable to that in rat liver. Rat apolipoprotein E expressed in transfected cells was secreted from the cells into the medium. MH - Amino Acid Sequence ; Animal ; Apolipoproteins E/*GENETICS ; Base Sequence ; Cloning, Molecular ; DNA/METABOLISM ; DNA Restriction Enzymes ; *Genes, Structural ; Human ; Introns ; L Cells/ENZYMOLOGY ; Liver/ *METABOLISM ; Mice ; Nucleic Acid Conformation ; Nucleoside Monophosphate Kinases/GENETICS ; Rats ; RNA, Messenger/GENETICS ; Sequence Homology, Nucleic Acid ; Support, Non-U.S. Gov't SO - J Biol Chem 1986 Oct 15;261(29):13777-83 16 UI - 87008540 AU - Haddad IA ; Ordovas JM ; Fitzpatrick T ; Karathanasis SK TI - Linkage, evolution, and expression of the rat apolipoprotein A-I, C-III, and A-IV genes. AB - The genes coding for three of the proteins of the lipid transport system, apolipoproteins A-I (apoA-I), C-III (apoC-III), and A-IV (apoA-IV), are closely linked and tandemly organized as a multigene family in the human genome. The evolution of this multigene family was studied by cloning and extensive restriction mapping analysis of an approximately 20-kilobase genomic DNA fragment containing the rat apoA-I gene. Low stringency hybridization blotting analysis of this DNA fragment using human apoC-III and apoA-IV cDNA probes revealed that the apoA-I, apoC-III, and apoA-IV genes are also closely linked and tandemly organized in the rat genome. Complete characterization of the rat apoA-I, apoC-III, and apoA-IV genes showed that their relative location, size, direction of transcription, and intron-exon organization are remarkably similar to those of the corresponding human genes. The relative steady state apoA-I, apoC-III, and apoA-IV mRNA levels in various rat tissues were determined by quantitative dot blot hybridization of tissue total RNA using the corresponding gene probes. Adult liver and intestine, but not colon, brain, spleen, muscle, heart, lung, and kidney, contain apoA-I, apoC-III, and apoA-IV mRNAs. Fetal liver and intestine contain apoA-I but not apoC-III or apoA-IV mRNAs. During neonatal development the liver contains apoA-I and apoC-III but not apoA-IV while the intestine contains apoA-I, apoC-III, and substantial amounts of apoA-IV mRNAs. In adulthood and during aging both liver and intestine contain all three apoA-I, apoC-III, and apoA-IV mRNAs. These results indicate that the apolipoprotein A-I/C-III/A-IV multigene family was established before mammalian radiation and suggest that these genes are similarly organized in the genomes of all mammals. In addition, these results indicate that expression of the rat apoA-I, apoC-III, and apoA-IV genes is liver- and intestine-specific and regulated by fetal-, neonatal-, and aging-related factors. MH - Amino Acid Sequence ; Animal ; Apolipoproteins A/*GENETICS ; Apolipoproteins C/*GENETICS ; Base Sequence ; DNA/ANALYSIS ; DNA Restriction Enzymes/METABOLISM ; *Gene Expression Regulation ; *Linkage (Genetics) ; Nucleic Acid Hybridization ; Rats ; RNA, Messenger/ANALYSIS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Tissue Distribution SO - J Biol Chem 1986 Oct 5;261(28):13268-77 17 UI - 87008488 AU - Chen SH ; Yang CY ; Chen PF ; Setzer D ; Tanimura M ; Li WH ; Gotto AM Jr ; Chan L TI - The complete cDNA and amino acid sequence of human apolipoprotein B-100. AB - We have determined the complete sequence of apolipoprotein (apo) B-100 cDNA. It is 14.1 kilobases in length and codes for a 4563-amino acid protein, including a 27-amino acid signal peptide and a 4536-amino acid mature protein. Further, we identified 2366 residues of apoB-100 by direct sequence analysis of apoB-100 tryptic peptides. The mature peptide is characterized by high hydrophobicity (0.916 kcal/residue) and predicted beta-sheet content (21%). Dot matrix analysis revealed the presence of many long internal repeats in apoB-100. The mature peptide contains 25 cysteine residues, 12 of which are in the N-terminal 500 residues. Twenty potential N-linked glycosylation sites were identified, of which 13 were proven to be glycosylated, and 4 were found not to be glycosylated by direct analysis of tryptic peptides. Our findings on apoB structure provide a basis for future experimentation on the role of apoB-100-containing lipoproteins in atherosclerosis. MH - Amino Acid Sequence ; Apolipoproteins B/ANALYSIS/*GENETICS ; Base Sequence ; Carbohydrates/ANALYSIS ; DNA/*ANALYSIS ; Human ; Hypercholesterolemia, Familial/BLOOD ; Peptide Fragments/METABOLISM ; Repetitive Sequences, Nucleic Acid ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Trypsin/METABOLISM SO - J Biol Chem 1986 Oct 5;261(28):12918-21 18 UI - 86229532 AU - Law A ; Wallis SC ; Powell LM ; Pease RJ ; Brunt H ; Priestley LM ; Knott TJ ; Scott J ; Altman DG ; Miller GJ ; et al TI - Common DNA polymorphism within coding sequence of apolipoprotein B gene associated with altered lipid levels. AB - 60 of 83 middle-aged white men had an XbaI restriction site polymorphism within the coding sequence of the apolipoprotein B gene. Subjects homozygous and heterozygous for the presence of an XbaI restriction site had mean serum triglyceride levels 36% higher (p = 0.02) than those in homozygotes without the restriction site; there was a less substantial difference (p = 0.03) in serum cholesterol. The findings supported a dominant pattern of inheritance. The presence of this restriction site may increase the risk of atherosclerotic disease. MH - Apolipoproteins B/*GENETICS ; Atherosclerosis/FAMILIAL & GENETIC ; Base Sequence ; Cholesterol/*BLOOD ; DNA/*ANALYSIS ; DNA Restriction Enzymes ; Homozygote ; Human ; Male ; Middle Age ; *Polymorphism (Genetics) ; Risk ; Triglycerides/*BLOOD SO - Lancet 1986 Jun 7;1(8493):1301-3 19 UI - 86296629 AU - Karathanasis SK ; Yunis I ; Zannis VI TI - Structure, evolution, and tissue-specific synthesis of human apolipoprotein AIV. AB - Apolipoprotein AIV (apoAIV) is a protein of the lipid transport system found associated with chylomicrons, high-density lipoprotein (HDL), and the lipoprotein-free fraction of the plasma. The gene coding for the human apoAIV is closely linked with the genes coding for apolipoproteins AI (apoAI) and CIII (apoCIII). In this paper a nearly full-length apoAIV cDNA clone has been isolated by screening an adult human liver DNA library using a human apoAIV gene probe. In-frame translation of the cDNA sequence in this clone indicated that the human apoAIV consists of 396 amino acid residues including a 20 residue long signal peptide. The coding region of this cDNA sequence contains 15 nucleotide repeats, 11 of which code for amino acid repeats with potentials of forming amphipathic helices. Alignment and comparison of the human and rat apoAIV amino acid sequences indicated a five-residue deletion near the carboxy terminus of the rat protein. This comparison also indicated that these proteins are 61.8% homologous, suggesting that the rate of evolution of apoAIV is 65 accepted point mutations (PAMs) per 100 residues per 100 million years. The rates of evolution of certain amino acid repeats in apoAIV are higher than the rate of evolution of the entire protein. However, the corresponding, computer-generated, secondary structures and hydropathy profiles of these repeats are very similar between the human and rat apoAIV. The relative steady-state levels of apoAIV mRNA in various human and monkey tissues were determined by hybridization blotting analysis of total RNA from these tissues using a human apoAIV cDNA probe.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Adult ; Amino Acid Sequence ; Animal ; Apolipoproteins A/BIOSYNTHESIS/ *GENETICS ; Base Sequence ; Cell Line ; Cells, Cultured ; Cloning, Molecular ; Comparative Study ; DNA/METABOLISM ; *Evolution ; Female ; Fetus ; *Genes, Structural ; Human ; Intestines/METABOLISM ; Liver/ METABOLISM ; Nucleic Acid Hybridization ; Pregnancy ; Protein Conformation ; Rats ; RNA, Messenger/GENETICS ; Species Specificity ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. ; Translation, Genetic SO - Biochemistry 1986 Jul 1;25(13):3962-70 20 UI - 86287319 AU - Protter AA ; Hardman DA ; Sato KY ; Schilling JW ; Yamanaka M ; Hort YJ ; Hjerrild KA ; Chen GC ; Kane JP TI - Analysis of cDNA clones encoding the entire B-26 region of human apolipoprotein B. AB - We report the characterization of intestine and liver cDNA clones for human apolipoprotein B (apoB) that map to the 5' end of the mRNA. The protein sequence encoded by the 5011 nucleotides derived from sequence analysis of these clones includes 1643 amino acid residues of the mature protein of Mr 184,000. The amino acid sequence at the amino terminus of B-74 peptide was determined and mapped to residue 1298. The size (Mr 145,700) and amino acid composition of the B-26 region encoded by these clones (including amino acid residues 1-1297) closely match the values obtained from the B-26 peptide. The amino acid sequence of peptide B-100 at the junction of peptides B-26 and B-74 (Phe-Lys decreases- Ser) shows structural homology to the site on human kininogen (Phe-Arg decreases- Ser) that is cleaved by the protease plasma kallikrein. The encoded protein contains five potential N-glycosylation sites and several regions in which the hydroxyamino acids, serine and threonine, are present in high abundance. The protein sequence presented in this report represents approximately 30% of the total B-100 protein and will aid in the characterization of additional cDNA clones. MH - Amino Acid Sequence ; Apolipoproteins B/*GENETICS/METABOLISM ; Base Sequence ; Cloning, Molecular ; DNA/GENETICS ; Human ; Intestines/ PHYSIOLOGY ; Liver/PHYSIOLOGY ; Protein Conformation ; Receptors, LDL/ METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Aug;83(15):5678-82 21 UI - 86286585 AU - Kunisada T ; Higuchi K ; Aota S ; Takeda T ; Yamagishi H TI - Molecular cloning and nucleotide sequence of cDNA for murine senile amyloid protein: nucleotide substitutions found in apolipoprotein A-II cDNA of senescence accelerated mouse (SAM). AB - cDNA clones encoding the murine senile amyloid protein (ASSAM) have been isolated from animal models of accelerated senescence (SAM-P/1) and from normal aging (SAM-R/1). Immunochemical and protein sequence studies revealed that apolipoprotein (apo) A-II is a serum precursor of ASSAM. A 17-base synthetic oligonucleotide based on residues 39-44 of ASSAM was used as a hybridization probe for screening newly constructed SAM-P/1 and SAM-R/1 liver cDNA libraries. The structure of murine apo A-II cDNA is of interest because of the amino acid substitution found in ASSAM and serum apo A-II of SAM-P; in SAM-R or other random bred slc:ICR mice, amino acid residue 5 of mature apo A-II is proline but, in SAM-P, this amino acid is changed to glutamine. This amino acid replacement is caused by two nucleotide substitutions (CCA for proline codon to CAG for glutamine codon). The third base mutation may not be relevant to the substitution of amino acid. Attention is directed to the relation of this amino acid substitution to the specific deposition of apo A-II, as a tissue amyloid fibril. MH - Aging ; Amino Acid Sequence ; Amyloid/*GENETICS ; Amyloid Protein SAA/ *GENETICS ; Animal ; Apolipoproteins/GENETICS ; Apolipoproteins A/ *GENETICS ; Base Sequence ; *Cloning, Molecular ; Comparative Study ; DNA/ ISOLATION & PURIFICATION/*METABOLISM ; DNA Restriction Enzymes ; *Genes, Structural ; Lipoproteins, HDL/*GENETICS ; Mice ; Mice, Mutant Strains ; Nucleic Acid Hybridization ; Poly A/ISOLATION & PURIFICATION ; RNA/ ISOLATION & PURIFICATION ; Species Specificity ; Support, Non-U.S. Gov't SO - Nucleic Acids Res 1986 Jul 25;14(14):5729-40 22 UI - 86284196 AU - Coleman RT ; Gonzalez PA ; Funke H ; Assmann G ; Levy-Wilson B ; Frossard PM TI - Polymorphisms in the apolipoprotein AI-CIII gene complex. AB - We report the presence of five restriction fragment length polymorphisms in the apolipoprotein AI-CIII (apo AI-CIII) gene complex as well as their respective allelic frequencies in a German population (pi) that we studied: an SstI (BanII) polymorphism in the 3' non-coding region of apo CIII q1(pi) = 0.13); an MspI polymorphism in the third intron of apo AI (q2(pi) = 0.12); a PvuII polymorphism in the first intron of apo CIII (q3(pi) = 0.30); and two XmnI polymorphisms in the 5' side of apo AI (q4(pi) = 0.20 and q5(pi) = 0.05). MH - Apolipoproteins A/*GENETICS ; Apolipoproteins C/*GENETICS ; Base Sequence ; DNA/*GENETICS ; DNA Restriction Enzymes ; Human ; Nucleic Acid Hybridization ; *Polymorphism (Genetics) ; Translation, Genetic SO - Mol Biol Med 1986 Jun;3(3):213-28 23 UI - 86256574 AU - Boguski MS ; Elshourbagy NA ; Taylor JM ; Gordon JI TI - Rat apolipoprotein A-IV: application of computational methods for studying the structure, function, and evolution of a protein. AB - There are a great variety of computational methods available to study protein and nucleic acid sequences. The choice of a computer program appropriate to a particular problem and the critical interpretation of the results can lead to specific, and experimentally testable, predictions of a protein's structure and function and may yield insights into its evolution and the location of its gene. We have shown that rat apoA-IV bears a striking structural similarity to human apoA-I (summarized in Fig. 7). Statistical analyses of homologies between apolipoproteins A-I, A-IV, and E demonstrate conclusively that all three sequences diverged from a common ancestral gene. That apoA-IV largely composed of 22 amino acid amphipathic segments with alpha-helical potential suggests that it possesses the structural requirements for LCAT activation. Analysis of the apoB, E receptor-binding domain of human apoE3 has demonstrated that it evolved from an ancestral repeated sequence. Assuming that the genes for these proteins evolved as a result of a series of intra- and intergenic unequal crossovers, it is likely that their genetic loci were at one time linked. The repeated sequences of which these genes are composed have propagated themselves in an expansionary manner. Given this fact, the existence of other genes or pseudogenes based upon this repeated sequence motif is a distinct possibility. MH - Amino Acid Sequence ; Animal ; Apolipoproteins A/*GENETICS ; Base Sequence ; Comparative Study ; *Evolution ; Human ; Information Systems ; Lipoproteins, HDL/GENETICS ; Protein Conformation ; Rats ; RNA, Messenger/ GENETICS ; Sequence Homology, Nucleic Acid ; Species Specificity ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Methods Enzymol 1986;128:753-73 24 UI - 86245590 AU - Funke H ; Rust S ; Assmann G TI - Detection of apolipoprotein E variants by an oligonucleotide "melting: procedure. AB - An oligonucleotide "melting: procedure was developed whereby we can monitor for a sequence heterogeneity in the gene for apolipoprotein E production. Two oligonucleotides were synthesized, one recognizing the common epsilon 3 allele but not the common epsilon 2 allele, the other recognizing the epsilon 2 allele but not the epsilon 3 allele. Samples from 15 subjects with different apolipoprotein E phenotypes as classified by isoelectric focusing were analyzed by this method for the presence of an Arg----Cys substitution in position 158 of the apolipoprotein E amino acid sequence. In 14 of these subjects the genotype determined by oligonucleotide melting agreed with the phenotype identified by isoelectric focusing. In one patient, however, whose phenotype was E2/2 by isoelectric focusing, the DNA hybridized with both oligonucleotides. We conclude that this patient has a mutation in one of his alleles for apolipoprotein E that differs from the frequently seen Arg----Cys change. The apolipoprotein E gene may thus be more heterogeneous than previously anticipated. MH - Alleles ; Apolipoproteins E/BLOOD/*GENETICS ; Base Sequence ; DNA/ BIOSYNTHESIS ; Electrophoresis, Agar Gel ; Human ; Hyperlipoproteinemia Type III/*FAMILIAL & GENETIC ; Isoelectric Focusing ; Isotope Labeling ; Leukocytes/METABOLISM ; Nucleic Acid Hybridization ; *Oligonucleotides/ CHEM SYNTHESIS ; Phenotype ; Support, Non-U.S. Gov't SO - Clin Chem 1986 Jul;32(7):1285-9 25 UI - 86082332 AU - Noteborn M ; Arnberg A ; de Jonge M ; Ab G ; Gruber M TI - Splicing pathways of the chicken apo very low density lipoprotein II (pre)messenger RNA. AB - The precursor-mRNA transcribed from the chicken apo very low density lipoprotein II gene was identified. This gene which is under full estrogen control and only expressed in the liver, possesses three introns. Splicing intermediates were characterized by hybridization with intron-specific probes, and by electron microscopy of R-loops. The introns appear to be excised in a non-obligatory order, but at different rates. MH - Animal ; Apolipoproteins/*GENETICS ; Base Sequence ; Chickens/*GENETICS ; DNA ; Electrophoresis, Agar Gel ; Lipoproteins, VLDL/*GENETICS ; Male ; Microscopy, Electron ; Nucleic Acid Hybridization ; Nucleic Acid Precursors/*METABOLISM ; RNA, Messenger/*METABOLISM ; Transcription, Genetic SO - FEBS Lett 1986 Jan 1;194(1):151-6 26 UI - 86242165 AU - Davison PJ ; Norton P ; Wallis SC ; Gill L ; Cook M ; Williamson R ; Humphries SE TI - There are two gene sequences for human apolipoprotein CI (apo CI) on chromosome 19, one of which is 4 kb from the gene for apo E. AB - A cDNA probe corresponding to the mRNA sequence for apolipoprotein E (apo E) was used to screen two independently-constructed human genomic libraries. Two recombinants (lambda E-2, and lambda E2-1), isolated using the apo E cDNA probe, also contain part or all of the apo CI gene. Hybridisation studies using both apo E and apo CI cDNA probes show that these two genes are in the same orientation and separated by 4 kb. MH - Apolipoproteins C/*GENETICS ; Apolipoproteins E/*GENETICS ; Base Sequence ; *Chromosomes, Human, 19-20 ; DNA/ANALYSIS ; DNA Restriction Enzymes/ METABOLISM ; Human ; Nucleic Acid Hybridization ; Polymorphism (Genetics) ; Support, Non-U.S. Gov't SO - Biochem Biophys Res Commun 1986 May 14;136(3):876-84 27 UI - 86196059 AU - Boguski MS ; Birkenmeier EH ; Elshourbagy NA ; Taylor JM ; Gordon JI TI - Evolution of the apolipoproteins. Structure of the rat apo-A-IV gene and its relationship to the human genes for apo-A-I, C-III, and E. AB - We have determined the nucleotide sequence of the rat apolipoprotein (apo-) A-IV gene and analyzed its structural and evolutionary relationships to the human apolipoprotein A-I, E, and C-III genes. The rat A-IV gene is 2.4 kilobases in size and consists of three exons (142, 126, and 1157 base pairs) interrupted by two introns (277 and 673 base pairs). The 5'-nontranslated region and most of the signal peptide are encoded by the first exon. Thus, the apo-A-IV gene lacks an intron in the 5'-nontranslated region of its mRNA in contrast to all other known apolipoprotein genes. Sequences coding for amphipathic docosapeptides span both the second and third exons of the rat A-IV gene. We demonstrate that this is also true for the human apolipoprotein genes. This gene family seems to have evolved by the duplication of an ancestral minigene that resulted in the formation of two exons. Thereafter, evolution of these sequences was dominated by intraexonic amplification of repeating units coding for amphipathic peptides. Sequence divergence of these repeats resulted in the functional differentiation of the apolipoproteins. However, conservation of the fundamental amphipathic pattern allowed members of this protein family to retain their lipid-binding properties. MH - Animal ; Apolipoproteins A/*GENETICS ; Apolipoproteins C/*GENETICS ; Apolipoproteins E/*GENETICS ; Base Sequence ; Cloning, Molecular ; DNA/ ANALYSIS ; DNA Restriction Enzymes/METABOLISM ; Human ; Models, Genetic ; *Nucleic Acid Conformation ; Rats ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Transcription, Genetic SO - J Biol Chem 1986 May 15;261(14):6398-407 28 UI - 86149325 AU - Protter AA ; Hardman DA ; Schilling JW ; Miller J ; Appleby V ; Chen GC ; Kirsher SW ; McEnroe G ; Kane JP TI - Isolation of a cDNA clone encoding the amino-terminal region of human apolipoprotein B. AB - A partial cDNA clone for the B-26 region of apolipoprotein B was isolated from an adult human liver DNA library by screening with an oligonucleotide probe derived from amino-terminal protein sequence obtained from purified B-26 peptide. Antisera against a synthetic 17-residue peptide whose amino acid sequence was encoded by the clone cross-reacts with apolipoproteins B-26, B-100, and B-48, but not with B-74. The nucleotide sequence immediately upstream from the amino terminus of B-26 codes for an apparent signal sequence, implying that the B-26 moiety is in an amino-terminal locus in the B-100 protein. That this sequence represents a 5' end region is further supported by primer extension analysis using a fragment of the cDNA clone and by S1 nuclease protection experiments using the corresponding region in a genomic clone. MH - Amino Acid Sequence ; Apolipoproteins B/*GENETICS/IMMUNOLOGY ; Base Sequence ; Chromosome Mapping ; Cloning, Molecular ; DNA/GENETICS ; Genes, Structural ; Human ; Immunosorbent Technics ; RNA, Messenger/ GENETICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Mar;83(5):1467-71 29 UI - 86111885 AU - Elshourbagy NA ; Walker DW ; Boguski MS ; Gordon JI ; Taylor JM TI - The nucleotide and derived amino acid sequence of human apolipoprotein A-IV mRNA and the close linkage of its gene to the genes of apolipoproteins A-I and C-III. AB - Both cDNA and genomic clones encoding human apolipoprotein (apo-) A-IV have been isolated and characterized. Southern blot analyses of apo-A-IV gene-containing cosmids revealed that the apo-A-IV gene is linked to the apo-A-I and apo-C-III genes within a 20-kilobase span of chromosome 11 DNA. The apo-A-IV gene is located about 14 kilobases downstream from the apo-A-I gene in the same orientation, with the apo-C-III gene located between them in the opposite orientation. The nucleotide sequence of the corresponding human apo-A-IV mRNA was determined, and the derived amino acid sequence showed that mature plasma apo-A-IV contained 376 residues. Throughout most of its length, human apo-A-IV was found to contain multiple tandem 22-residue repeated segments having amphipathic, alpha-helical potential. Amino acid substitutions within these homologous segments were generally conservative in nature. A comparison of the sequences of human and rat apo-A-IV revealed a 79% identity of amino acid positions in the amino-terminal 60 residues and a 58% identity in the remainder of the sequences, with the human protein containing 5 extra residues near the carboxyl terminus. An examination of the distribution of apo-A-IV mRNA in different tissues of the rat, marmoset, and man showed that apo-A-IV mRNA was abundant in both the liver and small intestine of the rat, but abundant in both the liver and small intestine of the marmoset and man. It was expressed in only trace amounts in all other tissues that were examined. These findings on the structure and expression of apo-A-IV and the close linkage of its gene to those of apo-A-I and apo-C-III suggest a regulatory relationship between the three genes. MH - Amino Acid Sequence ; Animal ; Apolipoproteins A/*GENETICS ; Apolipoproteins C/GENETICS ; Base Sequence ; Callithricidae ; Chromosomes, Human, 6-12 ; Comparative Study ; DNA/ANALYSIS ; Human ; Linkage (Genetics) ; RNA, Messenger/*ANALYSIS ; Sequence Homology, Nucleic Acid SO - J Biol Chem 1986 Feb 15;261(5):1998-2002 30 UI - 86200216 AU - Luo CC ; Li WH ; Moore MN ; Chan L TI - Structure and evolution of the apolipoprotein multigene family. AB - We present the complementary DNA and deduced amino acid sequence of rat apolipoprotein A-II (apoA-II), and the results of a detailed statistical analysis of the nucleotide and amino acid sequences of all the apolipoprotein gene sequences published to date: namely, those of human and rat apoA-I, apoA-II and apoE, rat apoA-IV, and human apoC-I, C-II and C-III. Our results indicate that the apolipoprotein genes have very similar genomic structures, each having a total of three introns at the same locations. Using the exon/intron junctions as reference points, we have obtained an alignment of the coding regions of all the genes studied. It appears that the mature peptide regions of these genes are almost completely made up of tandem repeats of 11 codons. The part of mature peptide region encoded by exon 3 contains a common block of 33 codons, whereas the part encoded by exon 4 contains a much more variable number of internal repeats of 11 codons. These genes have apparently evolved from a primordial gene through multiple partial (internal) and complete gene duplications. On the basis of the degree of homology of the various sequences, and the pattern of the internal repeats in these genes, we propose an evolutionary tree for the apolipoprotein genes and give rough estimates of the divergence times between these genes. Our results show that apoA-II has evolved extremely rapidly and that apoA-I and apoE also have evolved at high rates but some regions are better conserved than the others. The rate of evolution of individual regions seems to be related to the stringency of their functional requirements. MH - Amino Acid Sequence ; Animal ; Apolipoproteins/*GENETICS ; Apolipoproteins A/GENETICS ; Apolipoproteins C/GENETICS ; Apolipoproteins E/GENETICS ; Base Sequence ; Comparative Study ; DNA ; *Evolution ; *Genes ; Human ; Rats ; Repetitive Sequences, Nucleic Acid ; RNA, Messenger ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - J Mol Biol 1986 Feb 5;187(3):325-40 31 UI - 86082374 AU - Lorenzetti R ; Sidoli A ; Palomba R ; Monaco L ; Martineau D ; Lappi DA ; Soria M TI - Expression of the human apolipoprotein AI gene fused to the E. coli gene for beta-galactosidase. AB - The human apoAI gene was expressed in E. coli by in-frame fusion to a modified beta-galactosidase gene present in plasmid pUR291. The fused beta-galactosidase-apoAI gene product was expressed at a high level and was recognized by an anti-human apoAI antiserum. Besides the fused protein, at least one degradation product having an Mr similar to that of beta-galactosidase was present in high amounts in bacterial extracts. These results and those of a pulse-chase experiment indicate that degradation took place only in the apoAI moiety of the chimeric protein. MH - Apolipoproteins A/*GENETICS ; Base Sequence ; Beta-Galactosidases/ *GENETICS ; Cloning, Molecular ; DNA, Recombinant/METABOLISM ; Escherichia Coli/*GENETICS ; Galactosidases/*GENETICS ; Genes, Bacterial ; *Genes, Regulator ; Human ; Lipoproteins, HDL/*GENETICS ; Liver/ METABOLISM ; Plasmids SO - FEBS Lett 1986 Jan 6;194(2):343-6