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PMID: 2507523 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Sequence and transcription mapping of Bacillus subtilis competence genes comB and comA, one of which is related to a family of bacterial regulatory determinants.

Journal of bacteriology ·Vol. 171 ·No. 10 ·1989-10-00 ·Pages 5362-75

Weinrauch Y, Guillen N, Dubnau DA

Abstract

The complete nucleotide sequences of the comA and comB loci of Bacillus subtilis were determined. The products of these genes are required for the development of competence in B. subtilis and for the expression of late-expressing competence genes. The major 5' termini of both the comA and comB transcripts were determined. The inferred promoters of both comA and comB contained sequences that were similar to those found at the -10 and -35 regions of promoters that are used by sigma A-RNA polymerase, the primary form of this enzyme in vegetative cells. The comB gene was located approximately 3 kilobase pairs upstream of the comA gene and encoded a 409-amino-acid protein with a predicted molecular weight of 46,693. The comA locus contained two open reading frames (ORFs) and comB contained one ORF. The predicted amino acid sequence of the comA ORF1 gene product consisted of 214 amino acids, with an aggregate molecular weight of 24,132. The ORF1 product was required for competence, while ORF2, which was cotranscribed with ORF1 and encoded a predicted protein of 126 amino acids, was not. The predicted protein sequence of the comA ORF1 gene product was found to be similar to that of several members of the effector class of procaryotic signal transducers. The C-terminal portion of the predicted comA sequence contained a possible helix-turn-helix motif, which is characteristic of DNA-binding proteins. comA ORF1 was cloned on a multicopy plasmid and was shown to complement the competence-deficient phenotype caused by the comA124 insertion of Tn917lac. Also, the presence of comA ORF1 in multiple copies interfered with sporulation. Anti-peptide antibodies raised to the predicted product of comA ORF1 reacted strongly with a single protein band of about 24,000 daltons in immunoblots. The possible roles of multiple signal transduction systems in triggering the development of competence are discussed.

MeSH Terms
Amino Acid Sequence Antibodies, Bacterial/immunology Bacillus subtilis/genetics Bacterial Proteins/genetics Base Sequence Blotting, Western Chromosome Mapping DNA Mutational Analysis DNA, Bacterial/genetics DNA-Binding Proteins/genetics Genes, Bacterial Genes, Regulator Genetic Complementation Test Molecular Sequence Data Promoter Regions, Genetic Single-Strand Specific DNA and RNA Endonucleases Transcription Factors/genetics,immunology Transcription, Genetic
Chemicals
Antibodies, Bacterial Bacterial Proteins ComA protein, Bacteria DNA, Bacterial DNA-Binding Proteins Transcription Factors comB protein, Haemophilus influenzae Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Weinrauch Y
Department of Microbiology, Public Health Research Institute, New York, New York 10016.
Guillen N
Dubnau D A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.10.5362-5375.1989
Published
1989-10-00
Pages
5362-75
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210374
Subset
IM
Grants
NIAID NIH HHS · AI10311 · United States
NCRR NIH HHS · RRNA-02990 · United States
Databases
GENBANK
M22855, M22856, M29689, M29851
External Links
PubMed source DOI: 10.1128/jb.171.10.5362-5375.1989 PMC Full Text View journal details
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