Functional Dissection of the LysR-type CysB Transcriptional Regulator

Łochowska, Anna; Iwanicka-Nowicka, Roksana; Płochocka, Danuta; Hryniewicz, Monika M.

doi:10.1074/jbc.m007192200

Cited by 91 publications

(30 citation statements)

References 43 publications

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“…2) (28). Downstream of cysA is an open reading frame encoding a polypeptide that exhibits high homology to the two LysR-type regulators of sulfur assimilation in E. coli, CysB and Cbl (22,27,34,65).…”

Section: Resultsmentioning

confidence: 99%

Isolation and Characterization ofBurkholderia cenocepaciaMutants Deficient in Pyochelin Production: Pyochelin Biosynthesis Is Sensitive to Sulfur Availability

Farmer

Thomas

2004

J Bacteriol

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The opportunistic pathogen Burkholderia cenocepacia produces the yellow-green fluorescent siderophore, pyochelin. To isolate mutants which do not produce this siderophore, we mutagenized B. cenocepacia with the transposon mini-Tn5Tp. Two nonfluorescent mutants were identified which were unable to produce pyochelin. In both mutants, the transposon had integrated into a gene encoding an orthologue of CysW, a component of the sulfate/thiosulfate transporter. The cysW gene was located within a putative operon encoding other components of the transporter and a polypeptide exhibiting high homology to the LysR-type regulators CysB and Cbl. Sulfate uptake assays confirmed that both mutants were defective in sulfate transport. Growth in the presence of cysteine, but not methionine, restored the ability of the mutants to produce pyochelin, suggesting that the failure to produce the siderophore was the result of a depleted intracellular pool of cysteine, a biosynthetic precursor of pyochelin. Consistent with this, the wild-type strain did not produce pyochelin when grown in the presence of lower concentrations of sulfate that still supported efficient growth. We also showed that whereas methionine and certain organosulfonates can serve as sole sulfur sources for this bacterium, they do not facilitate pyochelin biosynthesis. These observations suggest that, under conditions of sulfur depletion, cysteine cannot be spared for production of pyochelin even under iron starvation conditions.

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Section: Resultsmentioning

confidence: 99%

Isolation and Characterization ofBurkholderia cenocepaciaMutants Deficient in Pyochelin Production: Pyochelin Biosynthesis Is Sensitive to Sulfur Availability

Farmer

Thomas

2004

J Bacteriol

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“…It was unexpected that all the LeuO truncated variants had dimerization defects, as they have an intact LH (which is involved in dimer formation) (28-30, 34, 35). The effect observed in the truncated protein lacking only 5 residues has not been observed for other LTTRs (40,44,45). This is clear evidence that the C-terminal RD-II and RD-I subdomains of LeuO are involved not only in tetramer formation but also in dimerization.…”

Section: Smentioning

confidence: 69%

“…3), and these residues have been proposed to contact the DNA major groove (29). Ser54 corresponds to Ser35 in AmpR, Ser33 in OxyR, and Ser38 in CysB and GcvA, and these residues are required for DNA binding but not oligomerization (27,(43)(44)(45). On the other hand, the P139A mutant retained some ability to activate and almost fully repress transcription ( Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The DBD is the most conserved region between LTTRs, and mutations in this domain affect DNA binding, activation, and repression. Furthermore, mutations in the DBDs of CysB, OxyR, and GcvA were not transdominant, and it has been suggested that they are affected in oligomer formation (43)(44)(45). The LH that is also conserved among LTTRs has a role in dimer formation, and the less conserved RD-I domain is involved in binding and a coinducer response (27)(28)(29)(30).…”

Section: Smentioning

confidence: 99%

“…The selection of deletions was based on some that have been analyzed in other LTTRs (9,31,36,40,(43)(44)(45). Thus, fragments of leuO encoding amino acid residues 1 to 309, 1 to 304, 1 to 299, 1 to 294, 1 to 284, and 1 to 214 (Fig.…”

Section: Smentioning

confidence: 99%

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The Salmonella enterica Serovar Typhi LeuO Global Regulator Forms Tetramers: Residues Involved in Oligomerization, DNA Binding, and Transcriptional Regulation

et al. 2014

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LeuO is a LysR-type transcriptional regulator (LTTR) that has been described to be a global regulator in Escherichia coli and Salmonella enterica, since it positively and negatively regulates the expression of genes involved in multiple biological processes. LeuO Together, these data demonstrate that not only the C terminus but also the DBD of LeuO is involved in oligomer formation; therefore, each LeuO domain appears to act synergistically to maintain its regulatory functions in Salmonella enterica serovar Typhi.

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A novel LysR‐type regulator negatively affects biosynthesis of the immunosuppressant brasilicardin

Wolański

Krawiec

Schwarz

et al. 2020

Engineering in Life Sciences

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Brasilicardin A (BraA) is a promising immunosuppressive compound produced naturally by the pathogenic bacterium Nocardia terpenica IFM 0406. Heterologous host expression of brasilicardin gene cluster showed to be efficient to bypass the safety issues, low production levels and lack of genetic tools related with the use of native producer. Further improvement of production yields requires better understanding of gene expression regulation within the BraA biosynthetic gene cluster (Bra‐BGC); however, the only so far known regulator of this gene cluster is Bra12. In this study, we discovered the protein LysRNt, a novel member of the LysR‐type transcriptional regulator family, as a regulator of the Bra‐BGC. Using in vitro approaches, we identified the gene promoters which are controlled by LysRNt within the Bra‐BGC. Corresponding genes encode enzymes involved in BraA biosynthesis as well as the key Bra‐BGC regulator Bra12. Importantly, we provide in vivo evidence that LysRNt negatively affects production of brasilicardin congeners in the heterologous host Amycolatopsis japonicum. Finally, we demonstrate that some of the pathway related metabolites, and their chemical analogs, can interact with LysRNt which in turn affects its DNA‐binding activity.

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Functional Dissection of the LysR-type CysB Transcriptional Regulator

Abstract: CysB is a tetrameric

Cited by 91 publications

References 43 publications

Isolation and Characterization ofBurkholderia cenocepaciaMutants Deficient in Pyochelin Production: Pyochelin Biosynthesis Is Sensitive to Sulfur Availability

Isolation and Characterization ofBurkholderia cenocepaciaMutants Deficient in Pyochelin Production: Pyochelin Biosynthesis Is Sensitive to Sulfur Availability

The Salmonella enterica Serovar Typhi LeuO Global Regulator Forms Tetramers: Residues Involved in Oligomerization, DNA Binding, and Transcriptional Regulation

A novel LysR‐type regulator negatively affects biosynthesis of the immunosuppressant brasilicardin

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