Proteus mirabilis ambient-temperature fimbriae: cloning and nucleotide sequence of the aft gene cluster
G Massad1, J F Fulkerson, D C Watson
1Division of Infectious Diseases, University of Maryland School of Medicine, Baltimore 21201, USA.
Abstract:
Uropathogenic Proteus mirabilis produces at least four types of fimbriae. Amino acid sequences from two peptides, derived by tryptic digestion of the structural subunit of one type of these fimbriae, the ambient-temperature fimbriae, were determined: NVVPGQPSSTQ and LIEGENQLNYNA. PCR primers, based on these sequences and that of the N terminus, were used to amplify a 359-bp fragment. A cosmid clone, isolated from a P. mirabilis genomic library by hybridization with the 359-bp PCR product, was used to determine the nucleotide sequence of the atf gene cluster. A 3,903-bp region encodes three polypeptides: AtfA, the structural subunit; AtfB, the chaperone; and AtfC, the outer membrane molecular usher. No fimbria-related genes are evident either 5' or 3' to the three contiguous genes. AtfA demonstrates significant amino acid sequence identity with type 1 major fimbrial subunits of several enteric species. The 359-bp PCR product hybridized strongly with all Proteus isolates (n = 9) and 25% of 355 Escherichia coli isolates but failed to hybridize with any of 26 isolates among nine other uropathogenic species. Ambient-temperature fimbriae of P. mirabilis may represent a novel type of fimbriae of enteric species.
Insights
Uropathogenic Proteus mirabilis ambient-temperature fimbriae may be a novel type. Researchers identified the atf gene cluster, encoding key fimbrial proteins, suggesting a new class of enteric fimbriae.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Uropathogenic Proteus mirabilis possesses multiple fimbrial types.
- Fimbriae play crucial roles in bacterial adhesion and pathogenesis.
Purpose of the Study:
- To characterize the ambient-temperature fimbriae of Proteus mirabilis.
- To identify and analyze the genetic basis of these fimbriae.
Main Methods:
- Peptide sequencing of fimbrial subunits.
- Polymerase Chain Reaction (PCR) for gene amplification.
- Cosmid cloning and DNA sequencing of the atf gene cluster.
Main Results:
- Determined amino acid sequences of two peptides from the structural subunit.
- Amplified a 359-bp fragment using specific PCR primers.
- Sequenced a 3,903-bp region encoding AtfA (subunit), AtfB (chaperone), and AtfC (usher).
- AtfA shares sequence identity with type 1 fimbrial subunits of other enteric bacteria.
- PCR product showed strong hybridization with Proteus isolates and some E. coli, but not other uropathogens.
Conclusions:
- The ambient-temperature fimbriae of P. mirabilis are encoded by the contiguous atf gene cluster.
- These fimbriae may represent a novel type within enteric species.
- Further research is warranted to understand their specific role in uropathogenesis.
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