Proteus mirabilis MR/P fimbriae: molecular cloning, expression, and nucleotide sequence of the major fimbrial subunit gene.
Proteus mirabilis, a cause of serious urinary tract infection and acute pyelonephritis, produces several putative virulence determinants, among them, fimbriae. Principally, two fimbrial types are produced by this species: mannose-resistant/Proteus-like (MR/P) fimbriae and mannose-resistant/Klebsiella-like (MR/K) fimbriae. To isolate MR/P fimbrial gene sequences, a P. mirabilis cosmid library was screened by immunoblotting and by hybridization with an oligonucleotide probe based on the N-terminal amino acid sequence of the isolated fimbrial polypeptide, ADQGHGTVKFVGSIIDAPCS. One clone, pMRP101, reacted strongly with a monoclonal antibody specific for MR/P fimbriae and with the DNA probe. This clone hemagglutinated both tannic acid-treated and untreated chicken erythrocytes with or without 50 mM D-mannose and was shown to be fimbriated by transmission electron microscopy. A 525-bp open reading frame, designated mrpA, predicted a 175-amino-acid polypeptide including a 23-amino-acid hydrophobic leader peptide. The unprocessed and processed polypeptides are predicted to be 17,909 and 15,689 Da, respectively. The N-terminal amino acid sequence of the processed fimbrial subunit exactly matched amino acid residues 24 to 43 predicted by the mrpA nucleotide sequence. The MrpA polypeptide shares 57% amino acid sequence identity with SmfA, the major fimbrial subunit of Serratia marcescens mannose-resistant fimbriae.
Bahrani
F K
FK
Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore 21201.
Mobley
H L
HL
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D010861:Fimbriae, Bacterial / Q000502:physiology* / Q000648:ultrastructure; D005798:Genes, Bacterial; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D046911:Macromolecular Substances; D008358:Mannose / Q000494:pharmacology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011513:Proteus mirabilis / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D011704:Pyelonephritis / Q000382:microbiology; D012091:Repetitive Sequences, Nucleic Acid; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D046911:Macromolecular Substances; C079120:mrpA protein, Proteus mirabilis; D008358:Mannose