RESUMO
The two-component system (TCS) is a conserved signal transduction module in bacteria. The Hik2-Rre1 system is responsible for transcriptional activation upon high-temperature shift as well as plastoquinone-related redox stress in the cyanobacterium Synechococcus elongatus PCC 7942. As heat-induced de novo protein synthesis was previously shown to be required to quench the heat-activated response, we investigated the underlying mechanism in this study. We found that the heat-inducible transcription activation was alleviated by the overexpression of dnaK2, which is an essential homolog of the highly conserved HSP70 chaperone and whose expression is induced under the control of the Hik2-Rre1 TCS. Phosphorylation of Rre1 correlated with transcription of the regulatory target hspA. The redox stress response was found to be similarly repressed by dnaK2 overexpression. Considered together with the previous information, we propose a negative feedback mechanism of the Hik2-Rre1-dependent stress response that maintains the cellular homeostasis mediated by DnaK2.
Assuntos
Proteínas de Bactérias , Synechococcus , Retroalimentação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Synechococcus/genética , Synechococcus/metabolismo , Resposta ao Choque Térmico , Proteínas de Choque Térmico HSP70/genética , Regulação Bacteriana da Expressão GênicaRESUMO
Cyanobacteria employ two-component sensor-response regulator systems to monitor and respond to environmental challenges. The response regulators RpaA, RpaB, Rre1 and RppA are integral to circadian clock function and abiotic stress acclimation in cyanobacteria. RpaA, RpaB and Rre1 are known to interact with ferredoxin or thioredoxin, raising the possibility of their thiol regulation. Here, we report that Synechocystis sp. PCC 6803 Rre1, RpaA and RpaB exist as higher-order oligomers under oxidising conditions and that reduced thioredoxin A converts them to monomers. We further show that these response regulators contain redox-responsive cysteine residues with an Em7 around -300 mV. These findings suggest a direct thiol modulation of the activity of these response regulators, independent of their cognate sensor kinases.
Assuntos
Synechocystis , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cianobactérias/genética , Cianobactérias/metabolismo , Regulação Bacteriana da Expressão Gênica/genética , Oxirredução , Compostos de Sulfidrila , Synechocystis/genética , Synechocystis/metabolismo , Tiorredoxinas/genética , Tiorredoxinas/metabolismoRESUMO
Two-component systems have been extensively described in the control of gene expression in response to different environmental signals in the cyanobacterium Synechocystis sp. PCC 6803. The Hik34-Rre1 two-component system has been shown to regulate a set of genes under certain stress conditions. Some evidence indicates that another histidine kinase, probably Hik2, is acting upstream of Rre1 in the regulation of some genes in response to hyperosmotic and salt stress. In the present study, a mis-annotation of the Rre1 protein has been identified and the correct version has been functionally characterized in vitro. By using EMSA assays, we have demonstrated that phosphorylation of Rre1 by Hik2 increases the affinity of the response regulator for the adhA promoter region, a gene that has been demonstrated previously to be specifically regulated by the Hik34-Rre1 system. These results suggest that Hik2 might cooperate with Hik34 in the regulation of the adhA gene by transferring the phosphoryl group to Rre1 under salt and hyperosmotic stress conditions.