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PMID: 8039900 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The Vibrio cholerae acfB colonization determinant encodes an inner membrane protein that is related to a family of signal-transducing proteins.

Infection and immunity ·Vol. 62 ·No. 8 ·1994-08-00 ·Pages 3289-98

Everiss KD, Hughes KJ, Kovach ME, Peterson KM

Abstract

Vibrio cholerae accessory colonization factor genes (acfA, B, C, and D) are required for efficient intestinal colonization. Expression of acf genes is under the control of a regulatory cascade that also directs the synthesis of cholera toxin and proteins involved in the biogenesis of the toxin-coregulated pilus. The gene for acfB was cloned by using an acfB::TnphoA fusion junction to probe a V. cholerae O395 bacteriophage lambda library. DNA sequence analysis revealed that acfB is predicted to encode a 626-amino-acid protein related to the V. cholerae HlyB and TcpI proteins. These three Vibrio proteins have amino acid sequence similarity in a region highly conserved among bacterial methyl-accepting chemotaxis proteins. Analysis of the predicted AcfB amino acid sequence suggests that this colonization determinant possesses a membrane topology and domain organization similar to those of methyl-accepting chemotaxis proteins. Heterologous expression of acfB in Escherichia coli generates four polypeptide species with apparent molecular masses of 34, 35, 74, and 75 kDa. The 74- and 75-kDa proteins appear to represent modified forms of the full-length AcfB protein. The 34- and 35-kDa polypeptide species most likely correspond to a C-terminal 274-amino-acid polypeptide that results from internal translation initiation of acfB mRNA. Localization studies with AcfB-PhoA hybrid proteins indicate that AcfB resides in the V. cholerae inner membrane. V. cholerae acfB::TnphoA mutants display an altered motility phenotype in semisolid agar. The relationship between AcfB and Vibrio motility and the amino acid similarities between AcfB and chemotaxis signal-transducing proteins suggest that AcfB may interact with the V. cholerae chemotaxis machinery. The data presented in this report provide preliminary evidence that acfB encodes an environmental sensor/signal-transducing protein involved in V. cholerae colonization.

Related Genes
acfB
MeSH Terms
Amino Acid Sequence Bacterial Proteins/analysis,chemistry,genetics Base Sequence Cloning, Molecular Conserved Sequence DNA, Bacterial/chemistry Genes, Bacterial Intestines/microbiology Membrane Proteins/analysis,genetics Molecular Sequence Data Recombinant Fusion Proteins/analysis Signal Transduction Vibrio cholerae/genetics,pathogenicity
Chemicals
AcfB protein, Vibrio cholerae Bacterial Proteins DNA, Bacterial Membrane Proteins Recombinant Fusion Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Everiss K D
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130.
Hughes K J
Kovach M E
Peterson K M
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Article Info
Journal
Infection and immunity
Abbr.
Infect Immun
ISSN
0019-9567
DOI
10.1128/iai.62.8.3289-3298.1994
Published
1994-08-00
Pages
3289-98
Language
English
Region
United States
NLM ID
0246127
PMCID
PMC302958
Subset
IM
Grants
NIAID NIH HHS · AI28502 · United States
Databases
GENBANK
L25660
External Links
PubMed source DOI: 10.1128/iai.62.8.3289-3298.1994 PMC Full Text View journal details
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