==================================BSR48================================== 48. Pilin/Pili/Fimbriae Especially structure, genes, regulation. 1 UI - 87123729 AU - Jacobson SH TI - P-fimbriated Escherichia coli in adults with renal scarring and pyelonephritis. AB - The commonest organism in urinary tract infections (UTI) is Escherichia coli. Pyelonephritogenic E.coli strains possess P-fimbriae which firmly attach to uroepithelial cells by recognition of a carbohydrate structure, alpha-D-Galp-(1-4)-beta-D-Galp, which is confined within all glycosphingolipids related to the human P-blood group antigens. Several investigators have studied virulence properties of E.coli and host resistance in relation to UTI. Uroepithelial cells from children and women with recurrent UTI have an increased capacity to bind E.coli. In contrast to previous studies the present one deals with patients with renal scarring, who constitute the major risk group among patients with UTI. P-fimbriae mediated binding to uroepithelial cells was studied and the risk of recurrent UTI in patients with renal scarring was determined. Ninety per cent of the E.coli isolates from female patients with acute non-obstructive pyelonephritis in this study possess P-fimbriae (I). The fecal E.coli colonies obtained from these patients were P-fimbriated in 55% compared to 11% of the fecal E.coli colonies from healthy controls. The P-blood group distribution in 56 female patients with renal scarring and a history of febrile UTI was the same as in a control group of 39 healthy subjects (II). A history of recurrent and/or early infections did not increase the percentage of the P1 blood group phenotype. Forty-nine female patients with renal scarring were prospectively investigated for the incidence of symptomatic UTI in relation to fecal colonization with P-fimbriated E.coli (III). Fifty-three per cent of the patients had altogether 65 episodes of symptomatic UTI during the three-year follow-up (0.036 infections per month). Eight patients (16%) had nine attacks of acute pyelonephritis and 4/5 of the tested E.coli strains from these patients were P-fimbriated. No relationship was demonstrated between the presence of P-fimbriated E.coli in the fecal flora and the development of subsequent acute pyelonephritis. The binding of P-fimbriated E.coli to uroepithelial cells from 19 female patients with renal scarring was studied with the fluorescence-activated cell sorting (FACS) analysis (IV). The uroepithelial cells from the patients with renal scarring exhibited a significantly higher binding capacity (p less than 0.01) than uroepithelial cells from healthy controls. Furthermore, uroepithelial cells from the patients with renal scarring and kidney insufficiency had a higher availability of P-fimbriae receptors on their uroepithelial cells than cells obtained from patients with renal scarring and normal renal function (r = -0.75, p less than 0.001) (V).(ABSTRACT TRUNCATED AT 400 WORDS) MH - Adolescence ; Adult ; *Bacterial Adhesion ; Bacteriuria ; Escherichia Coli Infections/FAMILIAL & GENETIC/*MICROBIOLOGY ; Feces/MICROBIOLOGY ; Female ; Glomerular Filtration Rate ; Human ; Kidney/*PATHOLOGY/ PHYSIOPATHOLOGY ; Middle Age ; P Blood-Group System ; Phenotype ; *Pili, Bacterial ; Pyelonephritis/FAMILIAL & GENETIC/*MICROBIOLOGY ; Recurrence ; Support, Non-U.S. Gov't ; Urinary Tract Infections/FAMILIAL & GENETIC/ *MICROBIOLOGY SO - Acta Med Scand [Suppl] 1986;713:1-64 2 UI - 87096235 AU - Przondo-Hessek A ; Ko HL ; Ciborowski P ; Roszkowski W ; Pulverer G TI - Opsonin independent interaction of Klebsiella strains with human polymorphonuclear leukocytes. AB - Nine encapsulated Klebsiella strains with different types of fimbriation and their nonencapsulated mutants were tested for their stimulatory potency for human polymorphonuclear leukocytes in the absence of opsonins. The luminol chemiluminescence assay was used for these experiments. It could be shown that the interaction between Klebsiella bacteria and human leukocytes is rather complex depending not only on the presence of capsules but also on the hydrophobicity of Klebsiella surface and on the type of fimbriation existing. MH - Human ; Klebsiella/GENETICS/*PHYSIOLOGY/ULTRASTRUCTURE ; Luminescence ; Mutation ; Neutrophils/*PHYSIOLOGY ; Pili, Bacterial/*PHYSIOLOGY ; Polysaccharides, Bacterial/*PHYSIOLOGY ; Support, Non-U.S. Gov't SO - Zentralbl Bakteriol Mikrobiol Hyg [A] 1986 Nov;262(4):522-30 3 UI - 87086716 AU - Hales BA ; Amyes SG TI - Acquisition of a gene encoding mannose-resistant haemagglutinating fimbriae by a resistance plasmid during long-term urinary infection. AB - Urine was collected twice weekly from one patient during a 25-month period. Escherichia coli harbouring a resistance plasmid, pUK28, which confers trimethoprim, ampicillin, sulphamethoxazole, spectinomycin and streptomycin resistance was identified in the urine. Carriage of strains containing plasmid pUK28 was observed during three separate periods which totalled 16 months, even though the patient did not receive antibacterial drug therapy for most of that time. The plasmid was able to acquire the genes responsible for mannose-resistant haemagglutination and these genes were increasingly associated with the plasmid towards the end of the study period. MH - Aged ; Antigens, Bacterial/ANALYSIS ; Conjugation, Genetic ; Escherichia Coli/*GENETICS/IMMUNOLOGY/ULTRASTRUCTURE ; Escherichia Coli Infections/ *MICROBIOLOGY ; Female ; *Genes, Bacterial ; Hemagglutination ; Human ; Mannose/PHARMACOLOGY ; *Pili, Bacterial/IMMUNOLOGY ; *R Factors ; Support, Non-U.S. Gov't ; Time Factors ; Urinary Tract Infections/ *MICROBIOLOGY SO - J Med Microbiol 1986 Dec;22(4):297-301 4 UI - 87085458 AU - Nagy LK ; Mackenzie T ; Pickard DJ ; Dougan G TI - Effects of immune colostrum on the expression of a K88 plasmid encoded determinant: role of plasmid stability and influence of phenotypic expression of K88 fimbriae. AB - Passaging of the K88-positive Escherichia coli strain CN6913 through synthetic medium containing immune colostrum gave rise to large numbers of K88-negative CN6913 variants. These K88-negative variants had all lost a single large plasmid known to encode the K88 genetic determinant. Four other large plasmids harboured by this strain were unaffected. Viable K88-positive and K88-negative variants of CN6913 accumulated at a similar rate in synthetic medium and in medium containing non-immune colostrum. In the presence of immune colostrum, viable cells of the K88-negative variant accumulated faster and to a greater extent in cultures than the K88-positive variant if incubated at 37 degrees C, which favours the phenotypic expression of K88. However, when similar cultures were incubated at 18 degrees C, a temperature known to inhibit phenotypic expression of K88, the accumulation of viable cells of the two variants was strictly comparable in all media and no loss of plasmid or increase in K88-negative variants was observed. Cells containing a pBR322-based K88-encoding recombinant plasmid were also eliminated by immune colostrum whereas cells containing pBR322 were not. Plasmids encoding the K99 antigen were not readily eliminated from strains passaged through medium containing immune colostrum. K99-negative variants that were detected still harboured the K99-encoding plasmid. MH - Animal ; Colostrum/*IMMUNOLOGY ; Escherichia Coli/GROWTH & DEVELOPMENT/ *IMMUNOLOGY ; Female ; Phenotype ; Pili, Bacterial/*IMMUNOLOGY ; *Plasmids ; Pregnancy ; Swine ; Temperature SO - J Gen Microbiol 1986 Sep;132 ( Pt 9):2497-503 5 UI - 87085456 AU - Tinsley CR ; Heckels JE TI - Variation in the expression of pili and outer membrane protein by Neisseria meningitidis during the course of meningococcal infection. AB - The occurrence of antigenic shift during meningococcal infection has been investigated by comparison of paired isolates obtained from the blood, cerebrospinal fluid or nasopharynx of patients. Isolates from any individual produced identical DNA 'fingerprints' and showed stability in expression of both class 2 outer membrane protein and an antigen common to pathogenic Neisseria, confirming their origin as a single strain. One of the four strains examined produced variants which differed in the molecular mass of their class 5 outer membrane proteins. Three of the strains produced pili containing the epitope recognized by monoclonal antibody SM1 and two of these gave rise to variants which expressed pili of differing subunit molecular masses. The two variants of the remaining strain produced pilins lacking the common epitope detected by antibody SM1 but radioimmune precipitation with polyclonal anti-pilus antiserum revealed that variation in the molecular mass of the pilin expressed also occurred with this second class of pili. Antigenic variation in expression of both class 5 outer membrane proteins and pili therefore appears to be a common occurrence during meningococcal infection. MH - Antigens, Bacterial/IMMUNOLOGY ; Antigens, Surface/IMMUNOLOGY ; Bacterial Outer Membrane Proteins/*IMMUNOLOGY ; Electrophoresis, Polyacrylamide Gel ; Human ; Meningitis, Meningococcal/*IMMUNOLOGY ; Neisseria Meningitidis/ *IMMUNOLOGY ; Pili, Bacterial/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; Variation (Genetics) SO - J Gen Microbiol 1986 Sep;132 ( Pt 9):2483-90 6 UI - 87085438 AU - Hales BA ; Amyes SG TI - The transfer of genes encoding production of mannose-resistant haemagglutinating fimbriae from uropathogenic enterobacteria. AB - Two hundred and thirty-seven bacterial strains, isolated from patients with urinary tract infections, were examined for the presence of plasmid-determined fimbrial adhesins. Ninety-nine strains were capable of producing mannose-resistant haemagglutination. Seventeen of these strains possessed transferable resistance plasmids and 11 of these were also able to transfer the mannose-resistant haemagglutination gene, suggesting that it was carried on the R plasmid or had been mobilized by it. MH - Bacterial Proteins/*GENETICS ; Bacteriuria/*MICROBIOLOGY ; DNA, Bacterial ; Enterobacteriaceae/*GENETICS ; Escherichia Coli/GENETICS ; *Genes, Bacterial ; Human ; Mannose/*GENETICS ; *Pili, Bacterial ; Proteus/ GENETICS ; R Factors ; Recombination, Genetic ; Support, Non-U.S. Gov't SO - J Gen Microbiol 1986 Aug;132 ( Pt 8):2243-7 7 UI - 87056058 AU - Ott M ; Hacker J ; Schmoll T ; Jarchau T ; Korhonen TK ; Goebel W TI - Analysis of the genetic determinants coding for the S-fimbrial adhesin (sfa) in different Escherichia coli strains causing meningitis or urinary tract infections. AB - Recently we have described the molecular cloning of the genetic determinant coding for the S-fimbrial adhesin (Sfa), a sialic acid-recognizing pilus frequently found among extraintestinal Escherichia coli isolates. Fimbriae from the resulting Sfa+ E. coli K-12 clone were isolated, and an Sfa-specific antiserum was prepared. Western blots indicate that S fimbriae isolated from different uropathogenic and meningitis-associated E. coli strains, including O83:K1 isolates, were serologically related. The Sfa-specific antibodies did not cross-react with P fimbriae, but did cross-react with F1C fimbriae. Furthermore the sfa+ recombinant DNAs and some cloned sfa-flanking regions were used as probes in Southern experiments. Chromosomal DNAs isolated from O18:K1 and O83:K1 meningitis strains with and without S fimbriae and from uropathogenic O6:K+ strains were hybridized against these sfa-specific probes. Only one copy of the sfa determinant was identified on the chromosome of these strains. No sfa-specific sequences were observed on the chromosome of E. coli K-12 strains and an O7:K1 isolate. With the exception of small alterations in the sfa-coding region the genetic determinants for S fimbriae were identical in uropathogenic O6:K+ and meningitis O18:K1 and O83:K1 strains. The sfa determinant was also detected on the chromosome of K1 isolates with an Sfa-negative phenotype, and specific cross-hybridization signals were visible after blotting against F1C-specific DNA. In addition homology among the different strains was observed in the sfa-flanking regions. MH - Antigens, Bacterial/GENETICS ; *Bacterial Adhesion ; Bacterial Proteins/ *GENETICS ; Chromosome Mapping ; DNA, Bacterial/GENETICS ; Escherichia Coli/*GENETICS/PATHOGENICITY ; Genes, Bacterial ; Human ; Meningitis/ *MICROBIOLOGY ; Molecular Weight ; *Pili, Bacterial ; Sequence Homology, Nucleic Acid ; Sialic Acids ; Species Specificity ; Support, Non-U.S. Gov't ; Urinary Tract Infections/*MICROBIOLOGY SO - Infect Immun 1986 Dec;54(3):646-53 8 UI - 87032398 AU - Korhonen TK ; Parkkinen J ; Hacker J ; Finne J ; Pere A ; Rhen M ; Holth:ofer H TI - Binding of Escherichia coli S fimbriae to human kidney epithelium. AB - Purified S fimbriae and an Escherichia coli strain carrying the recombinant plasmid pANN801-4 that encodes S fimbriae were tested for adhesion to frozen sections of human kidney. The fimbriae and the bacteria bound to the same tissue domains, and in both cases the binding was specifically inhibited by the receptor analog of S fimbria, sialyl(alpha 2-3)lactose. S fimbriae bound specifically to the epithelial elements in the kidneys; to the epithelial cells of proximal and distal tubules as well as of the collecting ducts and to the visceral and parietal glomerular epithelium. In addition, they bound to the vascular endothelium of glomeruli and of the renal interstitium. No binding to connective tissue elements was observed. The results suggest that the biological function of S fimbriae is to mediate the adhesion of E. coli to human epithelial and vascular endothelial cells. MH - Adenocarcinoma/PATHOLOGY ; *Bacterial Adhesion ; Epithelium/CYTOLOGY/ MICROBIOLOGY ; Escherichia Coli/GENETICS/*PHYSIOLOGY/ULTRASTRUCTURE ; Female ; Hemagglutination ; Human ; Kidney/*MICROBIOLOGY/PATHOLOGY ; Kidney Neoplasms/PATHOLOGY ; Male ; Pili, Bacterial/*PHYSIOLOGY/ ULTRASTRUCTURE ; Plasmids ; Support, Non-U.S. Gov't SO - Infect Immun 1986 Nov;54(2):322-7 9 UI - 87028218 AU - Swanson J ; Bergstr:om S ; Robbins K ; Barrera O ; Corwin D ; Koomey JM TI - Gene conversion involving the pilin structural gene correlates with pilus+ in equilibrium with pilus- changes in Neisseria gonorrhoeae. AB - Gonococci (Gc) exhibit pilus+----pilus- "phase transitions: at high frequency, but only some of the pilus- Gc can revert to pilus+ phenotype. We examined reversible phase transitions between pilus+ Gc and a particular pilus- variant (P-rp+ phenotype) whose pilin mRNA carries a unique block of nucleotides encoding an "assembly missense: pilin polypeptide. The results show that Gc pilus+ in equilibrium with P-rp+ transitions can result from intragenic recombination in which there is nonreciprocal exchange of partially homologous DNA sequences from a partial pilin gene (in silent, storage form) into the expression locus' complete pilin gene. Hence Gc pilus phase variation, like pilus antigenic variation, can occur by gene conversion of the pilin structural gene expression locus. MH - Bacterial Adhesion ; Bacterial Outer Membrane Proteins/*GENETICS ; Base Sequence ; DNA, Bacterial/GENETICS ; DNA, Recombinant ; Gene Conversion ; Gene Expression Regulation ; Genes, Bacterial ; Genes, Structural ; Neisseria Gonorrhoeae/*GENETICS ; Nucleic Acid Hybridization ; Pili, Bacterial/*PHYSIOLOGY ; RNA, Messenger/GENETICS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Translocation (Genetics) SO - Cell 1986 Oct 24;47(2):267-76 10 UI - 87008384 AU - Feutrier J ; Kay WW ; Trust TJ TI - Purification and characterization of fimbriae from Salmonella enteritidis. AB - A human isolate of Salmonella enteritidis which displayed strong pellicle formation during static broth culture and mannose-sensitive hemagglutination produced fimbriae which were morphologically indistinguishable from type 1 fimbriae of members of the family Enterobacteriaceae. Fimbrin was purified to homogeneity, and the apparent molecular weight (Mr, 14,400) was markedly lower than that reported for the type 1 fimbrin of Salmonella typhimurium (Mr, 22,100). This fimbrin contained 40% hydrophobic amino acids and lacked cysteine. The sequence of the N-terminal 64 amino acids was determined, and sequence alignment revealed that although the 18 N-terminal residues of the S. enteritidis molecule shared considerable homology with Escherichia coli and S. typhimurium type 1 fimbrins, the S. enteritidis fimbrin lacked a 6- to 9-residue terminal sequence present in the other type 1 fimbrins and, after residue 18, shared little homology with the E. coli sequence. Antibodies raised to the purified S. enteritidis fimbrin bound to surface-exposed conformational epitopes on the native fimbriae and displayed pronounced serospecificity. These antibodies were used in the isolation of a nonfimbriated Tn10 insertion mutant which was unable to hemagglutinate. MH - Amino Acid Sequence ; Antigens, Bacterial/IMMUNOLOGY ; Bacterial Proteins/ *ANALYSIS/GENETICS/IMMUNOLOGY/ISOLATION & PURIFICATION ; Cell Fractionation ; DNA Insertion Elements ; Hemagglutination ; Membrane Proteins/*ANALYSIS/GENETICS/IMMUNOLOGY/ISOLATION & PURIFICATION ; Molecular Weight ; Mutation ; Pili, Bacterial/*ANALYSIS ; Salmonella Enteritidis/ANALYSIS/GENETICS/*ULTRASTRUCTURE ; Support, Non-U.S. Gov't SO - J Bacteriol 1986 Oct;168(1):221-7 11 UI - 87005952 AU - Morrissey PM ; Dougan G TI - Expression of a cloned 987P adhesion-antigen fimbrial determinant in Escherichia coli K-12 strain HB101. AB - The properties of three independent enterotoxigenic Escherichia coli isolates known to express 987P adhesion fimbriae in a manner subject to phase variation were examined. Phase variation could not be correlated with any major changes in the plasmid DNA content of these strains or with readily detectable changes in any other tested phenotypic markers. The 987P genetic determinant from one of these strains, E. coli 987, was cloned into the non-fimbriated E. coli K-12 strains HB101, and expressed, using the cosmid vector system. 987P fimbriae produced by cells harbouring these recombinant plasmids (987P+ phenotype) could not be distinguished from 987P fimbriae produced by strain 987. Expression of 987P fimbriae from some recombinant plasmids was unstable but none of the recombinants exhibited the phase variation phenotype displayed by the parental strain. One recombinant plasmid, pPM200, contained an insert of strain 987 DNA of ca. 33 kb. The HB101[pPM200] displayed a rather stable 987P+ phenotype, but this was not true for several hosts, since pPM200 acquired approx. 20-kb deletions following transformations of E. coli K-12 strains other than HB101. The deletions mapped to the same region of pPM200 irrespective of the host strain transformed. Cells harbouring the deleted plasmids did not express 987P fimbriae (987P- phenotype). MH - *Cloning, Molecular ; Cosmids ; Escherichia Coli/*GENETICS/ULTRASTRUCTURE ; *Genes, Bacterial ; Microscopy, Electron ; Pili, Bacterial/ *ULTRASTRUCTURE ; Plasmids ; *Transcription, Genetic SO - Gene 1986;43(1-2):79-84 12 UI - 86310309 AU - de Graaf FK ; Klaasen P TI - Organization and expression of genes involved in the biosynthesis of 987P fimbriae. AB - A chromosomal DNA segment encoding the biosynthesis of 987P fimbriae was isolated by cosmid-cloning and subsequent subcloning into pBR322. The 12 kb DNA segment expressed five polypeptides with apparent molecular weights of 81,000, 39,000, 28,500, 20,500, and 16,500, respectively. The location of the corresponding genes was determined by insertional mutagenesis using Tn5. The 20.5 K polypeptide was identified as the 987P fimbrial subunit by its reaction with specific anti-987P antibodies. The 81, 39, and 28.5 K polypeptides appeared to be accessory proteins involved in 987P production. MH - Bacterial Proteins/GENETICS ; Cloning, Molecular ; Cosmids ; DNA Restriction Enzymes ; DNA, Bacterial/ISOLATION & PURIFICATION ; Escherichia Coli/*GENETICS/ULTRASTRUCTURE ; *Genes, Bacterial ; Genes, Structural ; Mutation ; Pili, Bacterial/*ULTRASTRUCTURE ; Plasmids SO - MGG 1986 Jul;204(1):75-81 13 UI - 86303039 AU - Keith BR ; Maurer L ; Spears PA ; Orndorff PE TI - Receptor-binding function of type 1 pili effects bladder colonization by a clinical isolate of Escherichia coli. AB - The role of type 1 pili in promoting bladder colonization was examined by constructing two mutant strains of a clinical Escherichia coli isolate. One mutant was isogenic to the parental strain save for a lesion in a gene required for pilus receptor binding; the other mutant was isogenic save for a lesion in the gene encoding the pilus structural subunit. Using mixed infections of the parental and mutant strains in an ascending rat cystitis model, we found that type 1-piliated mutants that lacked the receptor-binding function were as ineffective in bladder colonization as were mutants lacking the entire organelle. MH - Adhesiveness ; Animal ; Bacterial Vaccines/IMMUNOLOGY ; Bladder/ *MICROBIOLOGY ; Escherichia Coli/*PATHOGENICITY ; Human ; Mutation ; Pili, Bacterial/*PHYSIOLOGY ; Rats ; Receptors, Immunologic/*METABOLISM ; Support, U.S. Gov't, P.H.S. SO - Infect Immun 1986 Sep;53(3):693-6 14 UI - 86277927 AU - Worobec EA ; Frost LS ; Pieroni P ; Armstrong GD ; Hodges RS ; Parker JM ; Finlay BB ; Paranchych W TI - Location of the antigenic determinants of conjugative F-like pili. AB - The amino terminus of the pilin protein constitutes the major epitope of F-like conjugative pili studied to date (F, ColB2, R1-19, R100-1, and pED208). Anti-pED208 pilus antibodies were passed through a CNBr-Sepharose affinity column linked to bovine serum albumin which was conjugated to a synthetic peptide, AcP(1-12), containing the major epitope at the amino terminus of pED208 pilin. This allowed the separation of two classes of antibodies; one was specific for the amino terminus and bound to the column, while the other, which recognizes a second epitope on the pilus, did not bind to the column. In addition, antibodies were raised against two amino-terminal peptide-bovine serum albumin conjugates [AcP(1-8) and AcP(1-12)] to ensure a source of pure, high-titer antibodies directed against the amino terminus. The location of these antibodies on intact pili was assayed by immunoelectron microscopy with a protein A-gold technique. The amino terminus-specific antibodies did not bind to the sides of the pili but appeared to be associated with the pilus tip. In addition, these antibodies were found to bind to the vesicle-like structure at the base of the pilus. The anti-pilus antibodies not specific for the amino terminus (unbound immunoglobulin G) were found to bind to the sides of the pilus. Anti-F and anti-ColB2 pilus antibodies bound to the sides of F, ColB2, and R1-19 pili, which have only their secondary epitope in common.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Antibodies, Bacterial/IMMUNOLOGY ; Antibody Specificity ; Antigenic Determinants ; *Antigens, Bacterial ; Bacterial Outer Membrane Proteins/ *IMMUNOLOGY ; Colicin Factors ; *F Factor ; Gold/DIAGNOSTIC USE ; Microscopy, Electron ; Pili, Bacterial/*IMMUNOLOGY SO - J Bacteriol 1986 Aug;167(2):660-5 15 UI - 86274643 AU - Klemm P TI - Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli. AB - The expression of type 1 fimbriae in Escherichia coli is phase dependent, i.e. a cell is either completely fimbriated or bald. This phenomenon is due to the periodic inversion of a specific 300-bp DNA segment containing the promoter for the fimbrial subunit gene, fimA. The phase switch is controlled by the products of two regulatory genes, fimB and fimE, located upstream of fimA. The fimB and fimE proteins direct the phase switch into the 'on' and 'off' position, respectively. The DNA sequence of a 3000-bp region containing the two genes has been determined. The fimB and fimE proteins exhibit strong homology and have most likely originated by duplication of an ancestral gene. They are highly basic implying that they control the phase switch through interaction at the DNA level. MH - Amino Acid Sequence ; Bacterial Outer Membrane Proteins/GENETICS ; Base Sequence ; DNA, Recombinant/METABOLISM ; Escherichia Coli/*GENETICS ; *Genes, Bacterial ; *Genes, Regulator ; *Genes, Structural ; Pili, Bacterial/*PHYSIOLOGY ; Plasmids ; Support, Non-U.S. Gov't SO - EMBO J 1986 Jun;5(6):1389-93 16 UI - 86254275 AU - Old DC ; Crichton PB TI - P fimbriation among biochemically inactive strains of Escherichia coli of the group formerly called Alkalescens-Dispar. AB - Biochemically inactive, non-motile strains of Escherichia coli of the group formerly known as Alkalescens-Dispar (AD) and of known AD serogroups were analysed for biotype, resistotype, type-1 fimbriation, P fimbriation and haemolysin production. All strains of AD serogroups O1 and O2 examined were P-fimbriate and of closely related bio-resistotypes, suggesting that they may be representatives of two E. coli P-fimbriate clones, members of which have not infrequently been isolated from infected urine. MH - Drug Resistance, Microbial ; Escherichia Coli/*CLASSIFICATION/METABOLISM ; Hemagglutinins/ANALYSIS ; Hemolysins/BIOSYNTHESIS ; *Pili, Bacterial ; Serotyping SO - J Med Microbiol 1986 Jun;21(4):337-42 17 UI - 86251244 AU - de Ree JM ; Schwillens P ; van den Bosch JF TI - Monoclonal antibodies for serotyping the P fimbriae of uropathogenic Escherichia coli. AB - Monoclonal antibodies (MAbs) against seven serologically different P fimbriae (F7(1), F7(2), F8, F9, F11, F12, and F13) of uropathogenic Escherichia coli were tested for their ability to detect the P fimbriae on wild-type strains. In a plate agglutination test the MABs could detect the fimbriae on strains which expressed cloned fimbriae but not on wild-type strains. In a coagglutination test and in a whole-bacterium enzyme-linked immunosorbent assay the MAbs recognized the fimbriae on strains with cloned fimbriae and on wild-type strains. However, the coagglutination test has some disadvantages: only immunoglobulin G MAbs can be used, and the results cannot be read in an objective way. From these results, we concluded that the whole-bacterium enzyme-linked immunosorbent assay is the most convenient method for the determination of P fimbriae on wild-type E. coli strains. With this fast and easy method it is possible to do epidemiological studies on the distribution of P fimbriae among clinical isolates of uropathogenic E. coli and to extend the O:K:H serotype with the F serotype. MH - Agglutination Tests ; Antibodies, Monoclonal/*DIAGNOSTIC USE ; Comparative Study ; Enzyme-Linked Immunosorbent Assay ; Escherichia Coli/ *CLASSIFICATION/GENETICS/IMMUNOLOGY ; IgG/DIAGNOSTIC USE ; Pili, Bacterial/*IMMUNOLOGY ; Plasmids ; Support, Non-U.S. Gov't SO - J Clin Microbiol 1986 Jul;24(1):121-5 18 UI - 86250630 AU - Van Die I ; Van Megen I ; Zuidweg E ; Hoekstra W ; De Ree H ; Van den Bosch H ; Bergmans H TI - Functional relationship among the gene clusters encoding F7(1), F7(2), F9, and F11 fimbriae of human uropathogenic Escherichia coli. AB - The P fimbrial gene clusters encoding the serologically different F7(1), F7(2), F9, and F11 fimbriae were compared functionally. The results show that these gene clusters are closely related. MH - Bacterial Proteins/GENETICS ; Comparative Study ; Escherichia Coli/ *GENETICS/PATHOGENICITY/ULTRASTRUCTURE ; *Genes, Bacterial ; Human ; *Pili, Bacterial ; Urinary Tract/MICROBIOLOGY SO - J Bacteriol 1986 Jul;167(1):407-10 19 UI - 86250599 AU - Elleman TC ; Hoyne PA ; McKern NM ; Stewart DJ TI - Nucleotide sequence of the gene encoding the two-subunit pilin of Bacteroides nodosus 265. AB - The nucleotide sequence of the gene encoding pilin from Bacteroides nodosus 265 has been determined. The pilin is encoded by a single-copy gene, from which can be predicted a prepilin comprising a single protein chain of Mr 16,637. The prepilin sequence differs in several respects from the mature protein sequence. Seven additional N-terminal amino acid residues are present in prepilin, whereas residue 8, phenylalanine, undergoes posttranslational modification to become the N-methylated amino-terminal residue of mature pilin. In addition, further processing occurs through internal cleavage to produce two noncovalently linked subunits characteristic of pilins from serogroup H of B. nodosus, of which strain 265 is a member. The position of cleavage has been identified between alanine residues at positions 72 and 73 of the mature 149-residue pilin protein. The predicted pilin sequence of B. nodosus 265 shows extensive N-terminal amino acid sequence homology with other pilins of the N-methylphenylalanine type. In addition this sequence also shows homology with these N-methylphenylalanine-type pilins in the C-terminal region of the molecule, especially with pilin from Pseudomonas aeruginosa PAK. MH - Amino Acid Sequence ; Bacterial Outer Membrane Proteins/*GENETICS ; Bacteroides/*GENETICS ; Base Sequence ; Codon ; DNA, Bacterial/GENETICS ; Genes, Bacterial ; *Genes, Structural ; *Pili, Bacterial ; Protein Precursors/GENETICS SO - J Bacteriol 1986 Jul;167(1):243-50 20 UI - 86233338 AU - Bergstr:om S ; Robbins K ; Koomey JM ; Swanson J TI - Piliation control mechanisms in Neisseria gonorrhoeae. AB - Gonococci (Gc) undergo pilus+ to pilus- "phase transitions: readily in vitro. In the present study we sequenced pilin mRNA from reverting, pilus- Gc by oligonucleotide primer extension and compared these pilin mRNA sequences with those expressed by their pilus+ predecessors and pilus+ revertants. The results suggest that genetic rearrangement within the pilin structural gene can generate defective pilin gene products, resulting in a pilus- phenotype. These pilus- Gc give rise to pilus+ revertants upon reconstitution of their modified pilin gene. MH - Bacterial Outer Membrane Proteins/*GENETICS ; Base Sequence ; Cloning, Molecular ; DNA, Bacterial/GENETICS ; Genes, Bacterial ; Neisseria Gonorrhoeae/GENETICS/*PHYSIOLOGY ; Phenotype ; Pili, Bacterial/ *PHYSIOLOGY ; Protein Conformation ; RNA, Messenger/GENETICS ; Solubility ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Jun;83(11):3890-4 21 UI - 86226148 AU - Rhen M ; Tenhunen J ; V:ais:anen-Rhen V ; Pere A ; B~aga M ; Korhonen TK TI - Fimbriation and P-antigen recognition of Escherichia coli strains harbouring mutated recombinant plasmids encoding fimbrial adhesins of the uropathogenic E. coli strain KS71. AB - Deletion mutants of recombinant plasmids encoding the KS71B fimbrial antigens of the uropathogenic Escherichia coli strain KS71 (O4:K12) were constructed. The effects of these mutations were tested by transforming the mutated plasmids into non-fimbriated E. coli HB101 cells and testing the transformants for fimbriation and haemagglutination. A deletion transcriptionally upstream from the fimbrial subunit gene increased the expression of KS71B fimbriae. Deletion of the fimbrial subunit gene resulted in non-fimbriated but haemagglutinating transformants, whereas a deletion 6 kb transcriptionally downstream from the subunit gene resulted in non-haemagglutinating but fimbriate transformants, indicating that fimbriation and haemagglutination were genetically separable. We also present evidence suggesting that the fimbrillin and haemagglutinin are physically associated in the wild-type KS71 strain. MH - *Antigens, Bacterial ; Bacterial Proteins/*GENETICS ; DNA Restriction Enzymes ; *DNA, Recombinant ; Escherichia Coli/*GENETICS/IMMUNOLOGY ; Genes, Bacterial ; Hemagglutination ; Immunoglobulins, Fab/IMMUNOLOGY ; Mutation ; Pili, Bacterial/*IMMUNOLOGY ; *Plasmids ; Support, Non-U.S. Gov't SO - J Gen Microbiol 1986 Jan;132 ( Pt 1):71-7 22 UI - 86177566 AU - Segal E ; Hagblom P ; Seifert HS ; So M TI - Antigenic variation of gonococcal pilus involves assembly of separated silent gene segments. AB - The pilus is a major outer-membrane protein of Neisseria gonorrhoeae that undergoes phase and antigenic variation. In strain MS11 pilus expression is regulated at two expression loci on the chromosome, pilE1 and pilE2, although many other regions contain silent pilin information. A comparison of variant pilin sequences has revealed that the gene can be divided into a constant, a semivariable, and a hypervariable region. We report here that complete pilin genes are found only at the expression loci. Silent constant and variable region pilin gene segments are located on separate and distinct restriction fragments, and the generation of a complete pilin gene within the expression loci is the result of multiple recombination events. Conserved sequences within and flanking the pilin gene are proposed to act as recombination sites during the gene conversion events needed to produce a functional pilin gene. MH - Antigens, Bacterial/*GENETICS ; Bacterial Outer Membrane Proteins/ *GENETICS ; Gene Conversion ; Genes, Bacterial ; Neisseria Gonorrhoeae/ *GENETICS/IMMUNOLOGY ; Pili, Bacterial/*IMMUNOLOGY ; Signal Peptides/ GENETICS ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - Proc Natl Acad Sci USA 1986 Apr;83(7):2177-81 23 UI - 86176742 AU - Mooi FR ; Claassen I ; Bakker D ; Kuipers H ; de Graaf FK TI - Regulation and structure of an Escherichia coli gene coding for an outer membrane protein involved in export of K88ab fimbrial subunits. AB - The nucleotide sequence of the faeD gene of Escherichia coli and the amino acid sequence of its product is presented. The faeD product is an outer membrane protein required for transport of K88ab fimbrial subunits across the outer membrane. The protein is synthesized as a precursor containing a signal peptide, and the tentative mature protein comprises 777 amino acid residues. The distribution of amino acids in the faeD protein is similar to that of other outer membrane proteins; showing a fairly even distribution of charged residues and the absence of extensive hydrophobic stretches. Secondary structure predictions revealed a region of 250 amino acid residues which might be embedded in the outer membrane. The 5'-end of faeD is located within a region showing dyad symmetry. This region serves to couple translation of faeD to the translation of the gene preceding it (faeC). The 3'-end of faeD shows an overlap of 5 bases with the next gene (faeE). MH - Amino Acid Sequence ; Bacterial Outer Membrane Proteins/*GENETICS ; Bacterial Proteins/*GENETICS/METABOLISM ; Base Sequence ; Biological Transport ; DNA, Recombinant ; Escherichia Coli/*GENETICS ; Gene Expression Regulation ; Genes, Bacterial ; Nucleic Acid Conformation ; *Pili, Bacterial ; Protein Conformation ; RNA, Messenger/GENETICS ; Solubility ; Translation, Genetic SO - Nucleic Acids Res 1986 Mar 25;14(6):2443-57 24 UI - 86166841 AU - Dougan G ; Sellwood R ; Maskell D ; Sweeney K ; Liew FY ; Beesley J ; Hormaeche C TI - In vivo properties of a cloned K88 adherence antigen determinant. AB - An Escherichia coli strain of serotype O9:K36:H19 harboring the K88 recombinant plasmid pMK005 was able to efficiently colonize the small bowel of young piglets after oral infection. The strain expressed K88 antigen in vivo, and bacteria were detected in close association with the surface of the intestinal villi. Mice infected orally or intravenously with attenuated Salmonella typhimurium SL3261 harboring pMK005 were well protected against subsequent challenge with the highly virulent S. typhimurium SL1344. Anti-K88 antibodies were detected in the serum of mice immunized with SL3261(pMK005). MH - Animal ; Animals, Newborn ; Antigens, Bacterial/*GENETICS ; Bacterial Proteins/*GENETICS ; Bacterial Vaccines/IMMUNOLOGY ; Cloning, Molecular ; Diarrhea/*MICROBIOLOGY/PREVENTION & CONTROL ; Escherichia Coli/GENETICS/ IMMUNOLOGY/*PATHOGENICITY ; Gene Expression Regulation ; Intestines/ MICROBIOLOGY ; Mice ; Molecular Weight ; *Pili, Bacterial ; Salmonella Typhimurium/GENETICS/IMMUNOLOGY ; Swine ; Vaccines, Attenuated/IMMUNOLOGY SO - Infect Immun 1986 Apr;52(1):344-7 25 UI - 86149403 AU - Lindberg F ; Lund B ; Normark S TI - Gene products specifying adhesion of uropathogenic Escherichia coli are minor components of pili. AB - The papE, papF, and papG genes of uropathogenic Escherichia coli are dispensable for the synthesis and assembly of pili associated with pyelonephritis, called Pap pili. Phenotypically, papF and papG mediate digalactoside [alpha-D-Galp-(1----4)-beta-D-Galp)-specific adhesion. Although whole bacterial cells of a papE mutant bind to this receptor, purified pili from such a mutant do not. This is in contrast to pili purified from the wild type, which bind specifically. The DNA sequences of the papE and papF genes are presented, together with the deduced primary structure of the gene products. Both proteins have most of the features characteristic of Escherichia coli type 1 and Pap pilins. The PapE protein can be detected in the purified wild-type pilus by NaDodSO4/polyacrylamide gel electrophoresis followed by silver staining or by autoradiography of gels to which radioiodinated pili have been applied. In rabbits immunized with purified Pap pili, antibodies specific for both PapE and PapF are produced. We propose that PapE and PapF are minor pilins in the Pap pilus. MH - Amino Acid Sequence ; Antibodies, Bacterial/IMMUNOLOGY ; Bacterial Outer Membrane Proteins/*GENETICS/IMMUNOLOGY/ISOLATION & PURIFICATION ; Base Sequence ; Comparative Study ; DNA, Bacterial/GENETICS ; Escherichia Coli/ *GENETICS/PHYSIOLOGY/PATHOGENICITY ; Escherichia Coli Infections ; Human ; Pili, Bacterial/*ANALYSIS/PHYSIOLOGY ; Pyelonephritis/ETIOLOGY ; Sequence Homology, Nucleic Acid ; Support, Non-U.S. Gov't SO - Proc Natl Acad Sci USA 1986 Mar;83(6):1891-5 26 UI - 86139866 AU - Minion FC ; Abraham SN ; Beachey EH ; Goguen JD TI - The genetic determinant of adhesive function in type 1 fimbriae of Escherichia coli is distinct from the gene encoding the fimbrial subunit. AB - The role of type 1 fimbriae in the mannose-sensitive attachment of Escherichia coli to eucaryotic cells was investigated by deletion mutation analysis of a recombinant plasmid, pSH2, carrying the genetic information for the synthesis and expression of functional type 1 fimbriae. A mutant, pUT2002, containing a deletion remote from the structural gene encoding the 17-kilodalton subunit protein of type 1 fimbriae failed to agglutinate guinea pig erythrocytes even though the bacteria expressed fimbriae morphologically and antigenically indistinguishable from those produced by the intact recombinant plasmid. Fimbriae isolated from pUT2002 failed to agglutinate guinea pig erythrocytes, but reacted with a monoclonal antibody specific for quaternary structural determinants of type 1 fimbriae. Moreover, the dissociated fimbrial subunits from this mutant were indistinguishable from normal fimbriae by their migration during electrophoresis in sodium dodecyl sulfate-polyacrylamide gels, by their reactivity with a monoclonal antibody directed against a subunit-specific epitope, and in enzyme-linked immunosorbent assays with monospecific antisera. These results indicate that the adhesive functions in type 1 fimbriae are dependent on a factor(s) encoded by a gene other than those required for synthesis, assembly, and expression of the structural 17-kilodalton subunit. MH - Adhesiveness ; Antigenic Determinants ; Chromosome Deletion ; DNA, Recombinant ; Escherichia Coli/*GENETICS/METABOLISM/PHYSIOLOGY/ ULTRASTRUCTURE ; Genes, Bacterial ; Hemagglutination ; Mutation ; Pili, Bacterial/IMMUNOLOGY/*PHYSIOLOGY/ULTRASTRUCTURE ; Plasmids ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Bacteriol 1986 Mar;165(3):1033-6 27 UI - 86111634 AU - Finlay BB ; Pasloske BL ; Paranchych W TI - Expression of the Pseudomonas aeruginosa PAK pilin gene in Escherichia coli. AB - Pseudomonas aeruginosa is a piliated opportunistic pathogen. We have recently reported the cloning of the structural gene for the pilus protein, pilin, from P. aeruginosa PAK (B. L. Pasloske, B. B. Finlay, and W. Paranchych, FEBS Lett. 183:408-412, 1985), and in this paper we present evidence that this chimera (pBP001) expresses P. aeruginosa PAK pilin in Escherichia coli independent of a vector promoter. The strength of the promoter for the PAK pilin gene was assayed, and the cellular location of the pilin protein within E. coli was examined. This protein was present mainly in the inner membrane fraction both with and without its six-amino-acid leader sequence, but it was not assembled into pili. MH - Bacterial Outer Membrane Proteins/*GENETICS ; Cell Compartmentation ; Cell Membrane/METABOLISM ; Escherichia Coli/GENETICS ; Gene Expression Regulation ; *Pili, Bacterial ; Promoter Regions (Genetics) ; Protein Processing, Post-Translational ; Pseudomonas Aeruginosa/*GENETICS ; Support, Non-U.S. Gov't SO - J Bacteriol 1986 Feb;165(2):625-30 28 UI - 86111595 AU - Strom MS ; Lory S TI - Cloning and expression of the pilin gene of Pseudomonas aeruginosa PAK in Escherichia coli. AB - Many strains of Pseudomonas aeruginosa possess pili which have been implicated in the pathogenesis of the organism. This report presents the cloning and expression in Escherichia coli of the gene encoding the structural subunit of the pili of P. aeruginosa PAK. Total DNA from this strain was partially digested with Sau3A and inserted into the cloning vector pUC18. Recombinant E. coli clones were screened with oligonucleotide probes prepared from the constant region of the previously published amino acid sequence of the mature pilin subunit. Several positive clones were identified, and restriction maps were generated. Each clone contained an identical 1.1-kilobase HindIII fragment which hybridized to the oligonucleotide probes. Western blot analysis showed that all of the clones expressed small amounts of the P. aeruginosa pilin subunit, which has a molecular mass of ca. 18,000. This expression occurred independently of the orientation of the inserted DNA fragments in the cloning vector, indicating that synthesis was directed from an internal promoter. However, subclones containing the 1.1-kilobase HindIII fragment in a specific orientation produced an order of magnitude more of the pilin subunit. While the expressed pilin antigen was located in both the cytoplasmic and outer membrane fractions of E. coli, none appeared to be polymerized into a pilus structure. MH - Bacterial Outer Membrane Proteins/*GENETICS/METABOLISM ; Cell Compartmentation ; Cell Membrane ; Cloning, Molecular ; DNA Restriction Enzymes/DIAGNOSTIC USE ; Escherichia Coli/GENETICS ; Gene Expression Regulation ; Genes, Bacterial ; Genes, Structural ; *Pili, Bacterial ; Plasmids ; Pseudomonas Aeruginosa/*GENETICS ; Support, U.S. Gov't, P.H.S. SO - J Bacteriol 1986 Feb;165(2):367-72 29 UI - 86084433 AU - Elleman TC ; Hoyne PA ; Emery DL ; Stewart DJ ; Clark BL TI - Expression of the pilin gene from Bacteroides nodosus in Escherichia coli. AB - Bacterial plasmids that direct the expression in Escherichia coli of the pilin of Bacteroides nodosus were constructed. The quantity of pilin produced was greater than that of the pilin synthesized by B. nodosus, but no surface structural pili were present; pilin was found associated with the inner membrane of E. coli. Vaccination of sheep with E. coli containing pilin elicited increases in agglutinating and enzyme-linked immunosorbent assay antibody titers, which in turn were lower than the titers in sheep immunized with pilin from B. nodosus. The E. coli-produced pilin vaccine initially appeared to delay the progression of infection in immunized sheep after a challenge with virulent homologous B. nodosus, but at a later time the severity of foot rot was similar to that in sheep vaccinated with a placebo. MH - Animal ; Antigens, Bacterial/GENETICS/IMMUNOLOGY ; Bacterial Outer Membrane Proteins/*GENETICS ; Bacteroides/*IMMUNOLOGY ; Cloning, Molecular ; Escherichia Coli/*GENETICS ; Foot Rot/IMMUNOLOGY/MICROBIOLOGY ; Gene Expression Regulation ; Molecular Weight ; *Pili, Bacterial ; Plasmids ; Sheep ; Vaccination SO - Infect Immun 1986 Jan;51(1):187-92 30 UI - 86079567 AU - Haas R ; Meyer TF TI - The repertoire of silent pilus genes in Neisseria gonorrhoeae: evidence for gene conversion. AB - To investigate the significance of silent gene loci for pilus antigenic variation in N. gonorrhoeae, we determined the nucleotide sequence of the major silent locus, pilS1. The pilS1 locus contains six tandem pilus gene copies linked by a 39 bp repeat sequence also present in the expression loci. All silent copies lack the common N-terminal coding sequence of pilin, containing instead variant sequence information that constitutes a semivariable (SV) and a hypervariable (HV) domain. The SV and HV domains are interspersed with short, strictly conserved (C) regions flanking small cassettes of variable sequence information. It appears that such minicassettes from silent copies can be duplicated and transferred to other silent or expression genes by means of gene conversion. MH - Amino Acid Sequence ; Antigens, Bacterial/*GENETICS ; Base Sequence ; *Gene Conversion ; Genes, Bacterial ; Neisseria Gonorrhoeae/*GENETICS ; Pili, Bacterial/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; Variation (Genetics) SO - Cell 1986 Jan 17;44(1):107-15