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PMID: 11741847 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of the metC-cysK operon, involved in sulfur metabolism in Lactococcus lactis.

Journal of bacteriology ·Vol. 184 ·No. 1 ·2002-01-00 ·Pages 82-90

Fernández M, Kleerebezem M, Kuipers OP, Siezen RJ, van Kranenburg R

Abstract

Sulfur metabolism in gram-positive bacteria is poorly characterized. Information on the molecular mechanisms of regulation of genes involved in sulfur metabolism is limited, and no regulator genes have been identified. Here we describe the regulation of the lactococcal metC-cysK operon, encoding a cystathionine beta-lyase (metC) and cysteine synthase (cysK). Its expression was shown to be negatively affected by high concentrations of cysteine, methionine, and glutathione in the culture medium, while sulfur limitation resulted in a high level of expression. Other sulfur sources tested showed no significant effect on metC-cysK gene expression. In addition we found that O-acetyl-l-serine, the substrate of cysteine synthase, was an inducer of the metC-cysK operon. Using a random mutagenesis approach, we identified two genes, cmbR and cmbT, involved in regulation of metC-cysK expression. The cmbT gene is predicted to encode a transport protein, but its precise role in regulation remains unclear. Disruption of cmbT resulted in a two- to threefold reduction of metC-cysK transcription. A 5.7-kb region containing the cmbR gene was cloned and sequenced. The encoded CmbR protein is homologous to the LysR family of regulator proteins and is an activator of the metC-cysK operon. In analogy to CysB from Escherichia coli, we propose that CmbR requires acetylserine to be able to bind the activation sites and subsequently activate transcription of the metC-cysK operon.

MeSH Terms
Bacterial Proteins/genetics Carrier Proteins/genetics,metabolism Cloning, Molecular Cysteine Synthase/genetics Gene Expression Regulation, Bacterial Genes, Bacterial Genes, Regulator Lactococcus lactis/genetics,metabolism Lyases/genetics Membrane Transport Proteins Molecular Sequence Data Operon/genetics Physical Chromosome Mapping Sequence Analysis, DNA Transcription Factors/genetics,metabolism
Chemicals
Bacterial Proteins Carrier Proteins CmbR protein, Lactococcus lactis CmbT protein, Lactococcus lactis Membrane Transport Proteins Transcription Factors Cysteine Synthase Lyases cystathionine beta-lyase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Fernández María
Wageningen Centre for Food Sciences, Wageningen, and Department of Flavour, Nutrition and Ingredients, NIZO Food Research, Ede, The Netherlands.
Kleerebezem Michiel
Kuipers Oscar P
Siezen Roland J
van Kranenburg Richard
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.1.82-90.2002
Published
2002-01-00
Pages
82-90
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC134770
Subset
IM
Databases
GENBANK
AF240531
External Links
PubMed source DOI: 10.1128/JB.184.1.82-90.2002 PMC Full Text View journal details
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