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PMID: 2793826 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.

Characterization of Acinetobacter calcoaceticus catM, a repressor gene homologous in sequence to transcriptional activator genes.

Journal of bacteriology ·Vol. 171 ·No. 10 ·1989-10-00 ·Pages 5410-21

Neidle EL, Hartnett C, Ornston LN

Abstract

Two structural genes needed for catechol degradation, catA and catB, encode the respective enzymes catechol 1,2-dioxygenase (EC 1.13.11.1) and muconate cycloisomerase (EC 5.5.1.1). Catechol is an intermediate in benzoate degradation, and the catA and catB genes are clustered within a 17-kilobase-pair (kbp) region of Acinetobacter calcoaceticus chromosomal DNA containing all of the structural genes required for the conversion of benzoate to tricarboxylic acid cycle intermediates. catA and catB were transcribed in the same direction and were separated by 3.8 kbp of DNA. The 3.8-kbp sequence revealed that directly downstream from catA and potentially transcribed in the same direction were two open reading frames encoding polypeptides of 48 and 36 kilodaltons (kDa). Genetic disruption of these open reading frames did not discernably alter either catechol metabolism or its regulation. A third open reading frame, beginning 123 bp upstream from catB and transcribed divergently from this gene, was designated catM. This gene was found to encode a 28-kDa trans-acting repressor protein that, in the absence of cis,cis-muconate, prevented expression of the cat structural genes. Constitutive expression of the genes was caused by a mutation substituting Arg-156 with His-156 in the catM-encoded repressor. The repressor protein proved to be a member of a diverse family of procaryotic regulatory proteins which, with rare exception, are transcriptional activators. Repression mediated by catM was not the sole transcriptional control exercised over catA in A. calcoaceticus. Expression of catA was elicited by either benzoate or cis,cis-muconate in a genetic background from which catM had been deleted. This induction required DNA in a segment lying 1 kbp upstream from the catA gene. It is likely that an additional gene, lying outside the region containing the structural genes necessary for benzoate metabolism, contributes to this control.

MeSH Terms
Acinetobacter/genetics Amino Acid Sequence Bacterial Proteins Base Sequence Cloning, Molecular DNA, Bacterial/genetics DNA-Binding Proteins Genes, Bacterial Genes, Regulator Molecular Sequence Data Protein Biosynthesis Repressor Proteins/genetics Structure-Activity Relationship Transcription Factors/genetics Transcription, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins Repressor Proteins Transcription Factors catM protein, Acinetobacter
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Neidle E L
Department of Biology, Yale University, New Haven, Connecticut 06511.
Hartnett C
Ornston L N
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.10.5410-5421.1989
Published
1989-10-00
Pages
5410-21
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210378
Subset
IM
Grants
NIGMS NIH HHS · GM33377 · United States
Databases
GENBANK
M23245, M29714, M29848
External Links
PubMed source DOI: 10.1128/jb.171.10.5410-5421.1989 PMC Full Text View journal details
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