Functional analysis of the Bacillus subtilis cysK and cysJI genes

Ploeg, Jan van der

doi:10.1016/s0378-1097(01)00225-7

Cited by 26 publications

(33 citation statements)

References 12 publications

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“…3A). This initiation start site is slightly different from the one identified previously, which is located 46 bp upstream of the translation initiation codon of cysJ (37). The deduced Ϫ35 (TTTACT) and Ϫ10 (TAAAGT) boxes of the promoter are quite similar to the consensus sequences for A -dependent promoters.…”

Section: Resultscontrasting

confidence: 61%

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Identification of Bacillus subtilis CysL, a Regulator of the cysJI Operon, Which Encodes Sulfite Reductase

Guillouard¹,

Auger²,

Hullo³

et al. 2002

J Bacteriol

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The way in which the genes involved in cysteine biosynthesis are regulated is poorly characterized in Bacillus subtilis. We showed that CysL (formerly YwfK), a LysR-type transcriptional regulator, activates the transcription of the cysJI operon, which encodes sulfite reductase. We demonstrated that a cysL mutant and a cysJI mutant have similar phenotypes. Both are unable to grow using sulfate or sulfite as the sulfur source. The level of expression of the cysJI operon is higher in the presence of sulfate, sulfite, or thiosulfate than in the presence of cysteine. Conversely, the transcription of the cysH and cysK genes is not regulated by these sulfur sources. In the presence of thiosulfate, the expression of the cysJI operon was reduced 11-fold, whereas the expression of the cysH and cysK genes was increased, in a cysL mutant. A cis-acting DNA sequence located upstream of the transcriptional start site of the cysJI operon (positions ؊76 to ؊70) was shown to be necessary for sulfur source-and CysL-dependent regulation. CysL also negatively regulates its own transcription, a common characteristic of the LysR-type regulators. Gel mobility shift assays and DNase I footprint experiments showed that the CysL protein specifically binds to cysJ and cysL promoter regions. This is the first report of a regulator of some of the genes involved in cysteine biosynthesis in B. subtilis.All living organisms require sulfur for the synthesis of proteins and essential cofactors. Sulfur can be assimilated either from inorganic sources, such as sulfate and thiosulfate, or from organic sources, such as sulfate esters, sulfamates, and sulfonates. In Escherichia coli, sulfate is transported into the cell via an ATP-binding cassette-type sulfate-thiosulfate transport system (5, 11). Sulfate is subsequently reduced to sulfide by a series of enzymatic steps involving ATP sulfurylase, adenosine 5Ј-phosphosulfate kinase, 3Ј-phosphoadenosine 5Ј-phosphosulfate (PAPS) sulfotransferase, and sulfite reductase (Fig. 1). An O-acetyl-L-serine thiol-lyase condenses sulfide and O-acetylserine to form cysteine. In E. coli and Salmonella enterica serovar Typhimurium, at least 22 genes are required for the transport and reduction of sulfate and for its incorporation into cysteine. Most of these genes are coordinately regulated in the cysteine regulon (11). The high-level expression of these genes requires CysB, a LysR-type transcriptional activator, the inducer N-acetylserine, and sulfur-limiting conditions (11, 26). The CysB protein binds as a tetramer just upstream of the Ϫ35 promoter region of the positively regulated cys genes. The interaction of CysB with the inducer causes the activator to undergo a conformational change, allowing it to interact with the activation sites of the cysJ, cysK, and cysP promoters (7, 23). L-Cysteine, sulfide, and thiosulfate downregulate L-cysteine biosynthesis (11,12).In Bacillus subtilis, the assimilation of sulfate and the biosynthesis of cysteine may occur via similar pathways (Fig. 1). Indeed, the enzymes and the ...

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Section: Resultscontrasting

confidence: 61%

“…The cysH gene, which encodes PAPS sulfotransferase, is the first gene of an operon encoding a sulfate permease (CysP) and enzymes catalyzing the first steps of the sulfate assimilation pathway (18,19). The cysJ and cysI genes, encoding the two subunits of the sulfite reductase, were recently identified (37).…”

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confidence: 99%

Identification of Bacillus subtilis CysL, a Regulator of the cysJI Operon, Which Encodes Sulfite Reductase

Guillouard¹,

Auger²,

Hullo³

et al. 2002

J Bacteriol

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“…Sulfate is first transported into the cell via a sulfate permease, CysP, related to inorganic phosphate transporters (26). Sulfate is subsequently reduced to sulfide, probably in four steps involving the sequential action of ATP sulfurylase, adenosine 5Ј-phosphosulfate (APS) kinase, 3Ј-phosphoadenosine 5Ј-phosphosulfate (PAPS) reductase, and sulfite reductase (3,27,55). An O-acetylserine thiol-lyase, the cysK gene product, further condenses sulfide and O-acetylserine (OAS) to form cysteine (55).…”

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confidence: 99%

Global Control of Cysteine Metabolism by CymR inBacillus subtilis

et al. 2006

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YrzC has previously been identified as a repressor controlling ytmI expression via its regulation of YtlI activator synthesis in Bacillus subtilis. We identified YrzC as a master regulator of sulfur metabolism. Gene expression profiles of B. subtilis ⌬yrzC mutant and wild-type strains grown in minimal medium with sulfate as the sole sulfur source were compared. In the mutant, increased expression was observed for 24 genes previously identified as repressed in the presence of sulfate. Since several genes involved in the pathways leading to cysteine formation were found, we propose to rename YrzC CymR, for "cysteine metabolism repressor." A CymR-dependent binding to the promoter region of the ytlI, ssuB, tcyP, yrrT, yxeK, cysK, or ydbM gene was demonstrated using gel shift experiments. A potential CymR target site, TAAWNCN 2 ANTWNAN 3 ATMGGAA TTW, was found in the promoter region of these genes. In a DNase footprint experiment, the protected region in the ytlI promoter region contained this consensus sequence. Partial deletion or introduction of point mutations in this sequence confirmed its involvement in ytlI, yrrT, and yxeK regulation. The addition of O-acetylserine in gel shift experiments prevented CymR-dependent binding to DNA for all of the targets characterized. Transcriptome analysis of a ⌬cymR mutant and the wild-type strain also brought out significant changes in the expression level of a large set of genes related to stress response or to transition toward anaerobiosis.

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“…The genes involved in cysteine biosynthesis and sulfur assimilation in E. coli and Salmonella enterica serovar Typhimurium have been well characterized (reviewed in reference 37). More recently, cysteine biosynthesis has been studied in the gram-positive bacteria B. subtilis (24,63) and Lactococcus lactis (18), and also in the archaeon genus Methanosarcina (5,36). In contrast, cysteine biosynthesis and sulfur assimilation in the gram-positive Staphylococcus aureus have not been well studied.…”

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Role of a Cysteine Synthase in Staphylococcus aureus

et al. 2004

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The gram-positive human pathogen Staphylococcus aureus is often isolated with media containing potassium tellurite, to which it has a higher level of resistance than Escherichia coli. The S. aureus cysM gene was isolated in a screen for genes that would increase the level of tellurite resistance of E. coli DH5␣. The protein encoded by S. aureus cysM is sequentially and functionally homologous to the O-acetylserine (thiol)-lyase B family of cysteine synthase proteins. An S. aureus cysM knockout mutant grows poorly in cysteine-limiting conditions, and analysis of the thiol content in cell extracts showed that the cysM mutant produced significantly less cysteine than wild-type S. aureus SH1000. S. aureus SH1000 cannot use sulfate, sulfite, or sulfonates as the source of sulfur in cysteine biosynthesis, which is explained by the absence of genes required for the uptake and reduction of these compounds in the S. aureus genome. S. aureus SH1000, however, can utilize thiosulfate, sulfide, or glutathione as the sole source of sulfur. Mutation of cysM caused increased sensitivity of S. aureus to tellurite, hydrogen peroxide, acid, and diamide and also significantly reduced the ability of S. aureus to recover from starvation in amino acid-or phosphate-limiting conditions, indicating a role for cysteine in the S. aureus stress response and survival mechanisms.

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Functional analysis of the Bacillus subtilis cysK and cysJI genes

Cited by 26 publications

References 12 publications

Identification of Bacillus subtilis CysL, a Regulator of the cysJI Operon, Which Encodes Sulfite Reductase

Identification of Bacillus subtilis CysL, a Regulator of the cysJI Operon, Which Encodes Sulfite Reductase

Global Control of Cysteine Metabolism by CymR inBacillus subtilis

Role of a Cysteine Synthase in Staphylococcus aureus

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