Proteus mirabilis fimbriae: N-terminal amino acid sequence of a major fimbrial subunit and nucleotide sequences of the genes from two strains

Proteus mirabilis, a common cause of urinary tract infection in hospitalized and catheterized patients, produces mannose-resistant/klebsiella-like (MR/K) and mannose-resistant/proteus-like (MR/P) hemagglutinins. The gene encoding the major structural subunit of a fimbria, possibly MR/K, was identified in two strains. A degenerate oligonucleotide probe based on the N terminus of the Proteus uroepithelial cell adhesin and antiserum raised against the denatured polypeptide were used to screen a cosmid gene bank of strain HU1069. A cosmid clone that reacted with the probe and antiserum was identified, and a fimbria-like open reading frame was determined by nucleotide sequencing. The predicted N-terminal amino acid sequence of the processed polypeptide, ENETPAPKVSSTKGEIQLKG (residues 23 to 42), did not match the uroepithelial cell adhesin N terminus but, rather, matched exactly the N-terminal amino acid sequence of a polypeptide with an apparent molecular size of 19.5 kDa isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of a fimbrial preparation from strain HI4320 expressing MR/K hemagglutinin. By using an oligonucleotide from the HU1069 open reading frame, the fimbrial gene was isolated and sequenced from a cosmid gene bank clone of strain HI4320. A 552-bp open reading frame predicts a 184-amino-acid polypeptide including a 22-amino-acid hydrophobic leader sequence. The unprocessed polypeptide is predicted to be 18,921 Da; the processed polypeptide is predicted to be 16,749 Da. The predicted amino acid sequence of the polypeptide encoded by the gene, designated pmfA, displayed 36% exact matches with the mannose-resistant fimbrial subunit encoded by smfA of Serratia marcescens but only 15% exact matches with the predicted sequence encoded by mrkA of Klebsiella pneumoniae.

[1]  H. Mobley,et al.  Proteus mirabilis MR/P fimbriae: molecular cloning, expression, and nucleotide sequence of the major fimbrial subunit gene , 1993, Journal of bacteriology.

[2]  H. Mobley,et al.  Proteus mirabilis flagella and MR/P fimbriae: isolation, purification, N-terminal analysis, and serum antibody response following experimental urinary tract infection , 1991, Infection and immunity.

[3]  H. Holthöfer,et al.  Tissue-binding affinity of Proteus mirabilis fimbriae in the human urinary tract , 1990, Infection and immunity.

[4]  S. Clegg,et al.  Type V collagen as the target for type‐3 fimbriae, enterobacterial adherence organelles , 1990, Molecular microbiology.

[5]  H. Mobley,et al.  Hemagglutinin, urease, and hemolysin production by Proteus mirabilis from clinical sources. , 1990, The Journal of infectious diseases.

[6]  S. Clegg,et al.  Identification and characterization of the genes encoding the type 3 and type 1 fimbrial adhesins of Klebsiella pneumoniae , 1989, Journal of bacteriology.

[7]  Y. Nakabeppu,et al.  Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens , 1988, Journal of bacteriology.

[8]  S. Clegg,et al.  Molecular characterization of the type 3 (MR/K) fimbriae of Klebsiella pneumoniae , 1988, Journal of bacteriology.

[9]  S. Clegg,et al.  Cloning and characterization of the gene cluster encoding type 3 (MR/K) fimbriae of Klebsiella pneumoniae , 1988 .

[10]  J. Tenney,et al.  MR/K hemagglutination of Providencia stuartii correlates with adherence to catheters and with persistence in catheter-associated bacteriuria. , 1988, The Journal of infectious diseases.

[11]  H. Mobley,et al.  Urease-positive bacteriuria and obstruction of long-term urinary catheters , 1987, Journal of clinical microbiology.

[12]  R. Cook,et al.  Identification and characterization of a uroepithelial cell adhesin from a uropathogenic isolate of Proteus mirabilis , 1986, Infection and Immunity.

[13]  R. Adegbola,et al.  Antigenic relationships among type-3 fimbriae of Enterobacteriaceae revealed by immunoelectronmicroscopy. , 1985, Journal of medical microbiology.

[14]  U. Setia,et al.  Bacteremia in a long-term care facility. Spectrum and mortality. , 1984, Archives of internal medicine.

[15]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[16]  E. Tarkka,et al.  Type 3 fimbriae of Klebsiella sp.: molecular characterization and role in bacterial adhesion to plant roots , 1983, Journal of bacteriology.

[17]  W. Anthony,et al.  A prospective microbiologic study of bacteriuria in patients with chronic indwelling urethral catheters. , 1982, The Journal of infectious diseases.

[18]  R. Adegbola,et al.  Haemagglutinins and fimbriae of Morganella, Proteus and Providencia. , 1982, Journal of medical microbiology.

[19]  R. Doolittle,et al.  A simple method for displaying the hydropathic character of a protein. , 1982, Journal of molecular biology.

[20]  P. Larsson,et al.  Attachment of Proteus mirabilis to human urinary sediment epithelial cells in vitro is different from that of Escherichia coli , 1980, Infection and immunity.

[21]  H. Birnboim,et al.  A rapid alkaline extraction procedure for screening recombinant plasmid DNA. , 1979, Nucleic acids research.

[22]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[23]  B. Senior The special affinity of particular types of Proteus mirabilis for the urinary tract. , 1979, Journal of medical microbiology.

[24]  I. Ofek,et al.  Influence of pili on the virulence of Proteus mirabilis in experimental hematogenous pyelonephritis. , 1978, The Journal of infectious diseases.

[25]  D. Musher,et al.  Urease. The primary cause of infection-induced urinary stones. , 1976, Investigative urology.

[26]  P. Gower,et al.  The site of infection in 133 patients with bacteriuria. , 1973, Clinical nephrology.

[27]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[28]  K. Fairley,et al.  Simple test to determine the site of urinary-tract infection. , 1967, Lancet.

[29]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.