Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens

Mizunoe, Yoshimitsu; Nakabeppu, Yusaku; Sekiguchi, Mutsuo; Kawabata, Shun-ichiro; Moriya, Tsuneo; Amako, Kazunobu

doi:10.1128/jb.170.8.3567-3574.1988

Cited by 32 publications

(26 citation statements)

References 50 publications

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“…TAT GCC TCG CAA ACG GAT TTA ACC AAT CCT TGG CAA GAG CAG ATC The major fimbrial subunit, MrpA, most closely resembles the corresponding subunit of the mannose-resistant hemagglutinating fimbriae of S. marcescens (57% amino acid identity) (9,26). This identity is striking and has been the subject of a previous discussion (8).…”

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confidence: 70%

“…Both share about 50% amino acid identity with MrpI. The major structural subunit MrpA is most homologous with the major subunit (SmfA) of another mannose-resistant fimbria expressed by Serratia marcescens (26). The middle three gene products, MrpB, MrpC, and MrpD, are most homologous with corresponding pap gene products, PapH (4), PapC (30), and PapD (SwissProt accession number P15319), of the Galao(1-4)Gal-binding P fimbriae of uropathogenic E. coli.…”

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confidence: 96%

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Proteus mirabilis MR/P fimbrial operon: genetic organization, nucleotide sequence, and conditions for expression

Bahrani

Mobley

1994

J Bacteriol

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Proteus mirabilis, an agent of urinary tract infection, expresses at least four fimbrial types. Among these are the MR/P (mannose-resistant/Proteus-like) fimbriae. MrpA, the structural subunit, is optimally expressed at 37°C in Luria broth cultured statically for 48 h by each of seven strains examined. Genes encoding this fimbria were isolated, and the complete nucleotide sequence was determined. The mrp gene duster encoded by 7,293 bp predicts eight polypeptides: MrpI (22,133 Da), MrpA (17,909 Da), MrpB (19,632 Da), MrpC (96,823 Da), MrpD (27,886 Da), MrpE (19,470 Da), MrpF (17,363 Da), and MrpG (13,169 Da). mrpI is upstream of the gene encoding the major structural subunit gene mrpA and is transcribed in the direction opposite to that of the rest of the operon. All predicted polypeptides share -25% amino acid identity with at least one other enteric fimbrial gene product encoded by the pap, fim, smf, fan, or mrk gene clusters.Proteus mirabilis, commonly associated with nosocomially acquired urinary tract infection (12,24,29,37,45), expresses a number of proteins that may contribute to virulence, including urease (15), hemolysin (28,34,41), flagella (7,27,33), and fimbriae (1,7,32,38,39,45,46). Four fimbria-like structures have been purified from P. mirabilis strains for which the major structural subunits have been identified on sodium dodecyl sulfate (SDS)-polyacrylamide gels and subjected to N-terminal analysis (6-8, 20, 43, 46). The gene clusters for two of the fimbriae, MR/P (mannose-resistant/Proteus-like) fimbriae and PMF (P. mirabilis fimbriae), have been cloned, and the nucleotide sequences of the major subunit genes, mrpA (9) and pmfA (6), have been reported.Among the fimbriae of P. mirabilis, MR/P fimbriae are perhaps best understood. The presence of 7-to 8-nm-diameter, channelled fimbriae were first observed on bacterial cells capable of hemagglutinating erythrocytes in a mannose-resistant fashion (31). The MR/P hemagglutination pattern is expressed by virtually all strains (27,31) and elicits a serum immunoglobulin response following experimental ascending urinary tract infection in mice (7, 20). Here we report the complete nucleotide sequence for the mrp operon, the distribution of the genes among Proteus and other genera, and the conditions under which MrpA is expressed in the wild-type isolate.We have previously demonstrated that mice, experimentally infected with P. mirabilis, developed a serum immunoglobulin G response to MrpA, the major subunit of MR/P fimbriae, indicating that the fimbriae are expressed in the urinary tract (7). We sought to mimic these parameters by determining the optimal growth conditions for the in vitro expression of MR/P fimbriae. Expression of MrpA by P. mirabilis H14320, isolated from the urine of a woman with catheter-associated bacteriuria (16, 45), was assessed by Western blotting (immunoblotting) using rabbit polyclonal antiserum raised against purified MR/P fimbriae (Fig. 1A) Under these optimal conditions (Luria broth, 37°C, 48 h, static), expression of MrpA by...

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confidence: 70%

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confidence: 96%

Proteus mirabilis MR/P fimbrial operon: genetic organization, nucleotide sequence, and conditions for expression

Bahrani

Mobley

1994

J Bacteriol

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“…Based on these observations, we hypothesized that S. marcescens OxyR positively regulates fimbrial expression, resulting in reduced biofilm formation in the oxyR mutant strains. Fimbria activity is commonly measured through agglutination of eukaryotic cells, such as S. cerevisiae cells or erythrocytes (24,37,38). The oxyR-1 mutant was found to be severely deficient in the ability to agglutinate budding yeast cells (Fig.…”

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confidence: 99%

ASerratia marcescensOxyR Homolog Mediates Surface Attachment and Biofilm Formation

Shanks

Stella²,

Kalivoda³

et al. 2007

J Bacteriol

104

131

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OxyR is a conserved bacterial transcription factor with a regulatory role in oxidative stress response. From a genetic screen for genes that modulate biofilm formation in the opportunistic pathogen Serratia marcescens, mutations in an oxyR homolog and predicted fimbria structural genes were identified. S. marcescens oxyR mutants were severely impaired in biofilm formation, in contrast to the hyperbiofilm phenotype exhibited by oxyR mutants of Escherichia coli and Burkholderia pseudomallei. Further analysis revealed that OxyR plays a role in the primary attachment of cells to a surface. Similar to what is observed in other bacterial species, S. marcescens OxyR is required for oxidative stress resistance. Mutations in oxyR and type I fimbrial genes resulted in severe defects in fimbria-associated phenotypes, revealing roles in cell-cell and cell-biotic surface interactions. Transmission electron microscopy revealed the absence of fimbria-like surface structures on an OxyR-deficient strain and an enhanced fimbrial phenotype in strains bearing oxyR on a multicopy plasmid. The hyperfimbriated phenotype conferred by the multicopy oxyR plasmid was absent in a type I fimbrial mutant background. Real-time reverse transcriptase PCR indicated an absence of transcripts from a fimbrial operon in an oxyR mutant that were present in the wild type and a complemented oxyR mutant strain. Lastly, chromosomal P lac -mediated expression of fimABCD was sufficient to restore wild-type levels of yeast agglutination and biofilm formation to an oxyR mutant. Together, these data support a model in which OxyR contributes to early stages of S. marcescens biofilm formation by influencing fimbrial gene expression.

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“…1). (J. G. Kusters, Department of Medical Microbiology, Free University, Amsterdam, The Netherlands, generously constructed the tree.) For comparison with PapA, the two most closely related known pilin genes from other members of the family Enterobacteriaceae, i.e., MR/P of Proteus mirabilis (5,6) and SMF of Serratia marcescens (58), were included in the tree (7) (Fig. 1).…”

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Analysis of the F Antigen-Specific papA Alleles of Extraintestinal Pathogenic Escherichia coli Using a Novel Multiplex PCR-Based Assay

et al. 2000

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Polymorphisms in PapA, the major structural subunit and antigenic determinant of P fimbriae of extraintestinal pathogenic Escherichia coli, are of considerable epidemiological, phylogenetic, and immunotherapeutic importance. However, to date, no method other than DNA sequencing has been generally available for their detection. In the present study, we developed and rigorously validated a novel PCR-based assay for the 11 recognized variants of papA and then used the new assay to assess the prevalence, phylogenetic distribution, and bacteriological associations of the papA alleles among 75 E. coli isolates from patients with urosepsis. In comparison with conventional F serotyping, the assay was extremely sensitive and specific, evidence that papA sequences are highly conserved within each of the traditionally recognized F serotypes despite the diversity observed among F types. In certain strains, the assay detected serologically occult copies of papA, of which some were shown to represent false-negative serological results and others were shown to represent the presence of nonfunctional pap fragments. Among the urosepsis isolates, the assay revealed considerable segregation of papA alleles according to O:K:H serotype, consistent with vertical transmission within clones, but with exceptions which strongly suggested horizontal transfer of papA alleles between lineages. Sequencing of papA from two strains that were papA positive by probe and PCR but F negative in the new PCR assay led to the discovery of two novel papA variants, one of which was actually more prevalent among the urosepsis isolates than were several of the known papA alleles. These findings provide novel insights into the papA alleles of extraintestinal pathogenic E. coli and indicate that the F PCR assay represents a versatile new molecular tool for epidemiological and phylogenetic investigations which should make rapid, specific detection of papA alleles available to any laboratory with PCR capability.

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Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens

Cited by 32 publications

References 50 publications

Proteus mirabilis MR/P fimbrial operon: genetic organization, nucleotide sequence, and conditions for expression

Proteus mirabilis MR/P fimbrial operon: genetic organization, nucleotide sequence, and conditions for expression

ASerratia marcescensOxyR Homolog Mediates Surface Attachment and Biofilm Formation

Analysis of the F Antigen-Specific papA Alleles of Extraintestinal Pathogenic Escherichia coli Using a Novel Multiplex PCR-Based Assay

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