Comparative sequence analysis of the plasmid-encoded regulator of enteropathogenic Escherichia coli strains

• Published in: Infection and immunity 2001-09, Vol.69 (9), p.5553-5564
• Main Authors: OKEKE, Iruka N, BORNEMAN, Jade A, SHIN, Sooan, MELLIES, Jay L, QUINN, Laura E, KAPER, James B
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Base Sequence; bfpA gene; Biological and medical sciences; bundlin; Cell Line; Electrophoresis - methods; Escherichia coli; Escherichia coli - genetics; Escherichia coli - pathogenicity; Escherichia coli Infections - microbiology; Escherichia coli Proteins; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Bacterial - genetics; Genetic Variation; Genetics; Humans; insertion sequence IS1294; Microbiology; Molecular Genomics; Molecular Sequence Data; Operon; Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains; perA gene; perABC operon; perB gene; perC gene; Plasmids - genetics; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Promoter Regions, Genetic - genetics; Repressor Proteins - genetics; Repressor Proteins - metabolism; Sequence Analysis, DNA; Virulence - genetics
• ISSN: 0019-9567; 1098-5522
• DOI: 10.1128/IAI.69.9.5553-5564.2001

Enteropathogenic Escherichia coli (EPEC) strains that carry the EPEC adherence factor (EAF) plasmid were screened for the presence of different EAF sequences, including those of the plasmid-encoded regulator (per). Considerable variation in gene content of EAF plasmids from different strains was seen. However, bfpA, the gene encoding the structural subunit for the bundle-forming pilus, bundlin, and per genes were found in 96.8% of strains. Sequence analysis of the per operon and its promoter region from 15 representative strains revealed that it is highly conserved. Most of the variation occurs in the 5' two-thirds of the perA gene. In contrast, the C-terminal portion of the predicted PerA protein that contains the DNA-binding helix-turn-helix motif is 100% conserved in all strains that possess a full-length gene. In a minority of strains including the O119:H2 and canine isolates and in a subset of O128:H2 and O142:H6 strains, frameshift mutations in perA leading to premature truncation and consequent inactivation of the gene were identified. Cloned perA, -B, and -C genes from these strains, unlike those from strains with a functional operon, failed to activate the LEE1 operon and bfpA transcriptional fusions or to complement a per mutant in reference strain E2348/69. Furthermore, O119, O128, and canine strains that carry inactive per operons were deficient in virulence protein expression. The context in which the perABC operon occurs on the EAF plasmid varies. The sequence upstream of the per promoter region in EPEC reference strains E2348/69 and B171-8 was present in strains belonging to most serogroups. In a subset of O119:H2, O128:H2, and O142:H6 strains and in the canine isolate, this sequence was replaced by an IS1294-homologous sequence.