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SANTOS, J. S. Identification of transcription factors and cellular signals that regulate cspC gene expression from Caulobacter crescentus. 89 f. 2011. Masters thesis (Microbiology) Instituto de Ciencias Biomedicas, Universidade de Sao Paulo, Sao Paulo, 2011. The cold shock proteins belong to a family of proteins presenting a highly conserved domain, called cold shock domain (CSD). They are involved in various cellular processes, including adaptation to low temperature, nutritional stress, cell growth and stationary phase. In C. crescentus, a non-pathogenic α-proteobacteria, the cold shock proteins CspC and CspD have two CSDs and they are induced during stationary phase. This study aims to determine the transcription factors and cellular signals involved in cspC gene regulation in C. crescentus. In the present study we scanned a library of 4000 mutant clones with the Tn5 transposon, from which seven mutants were identified presenting reduced expression of cspC: CCNA03569 (hypothetical protein); CCNA02510 (polysaccharide deacetylase); CCNA01594 (ribosomal protein L11 methyltransferase); CCNA02186 (acetolactate synthase 3 regulatory subunit); CCNA00084 (bifunctional phosphoribosylaminoimidazolecarboxamide formyltransferase/IMP cyclohydrolase); CCNA03616 (sulfite reductase (NADPH)) e CCNA01448 (fructose 1,6bisphosphatase II). Expression assays in the presence of a conditioned medium, showed that cspC and cspD are apparently not induced in response to increased cell density. The phenotype of the mutant cspC was evaluated as to oxidative stress resistance in the stationary phase. The results showed that the ∆cspC strain is highly sensitive to hydrogen peroxide and superoxide but is not sensitive to organic hydroperoxide. The absence of cspC probably is compensated by dps, since the expression of this gene is increased in ∆cspC strain. In contrast, the transcritpion of katG is decreased, but the activities of KatG and SodB are not affected in the cspC mutant. Under conditions of stress caused by hydrogen peroxide, sucrose or salt, cspC expression is not affected. The sigma factors sigmas SigT and SigU and transcription regulator Fur are not involved in the regulation of cspC, but cspC expression is increased in ΔsigJ and decreased in ∆oxyR strains, respectively. It was also determined that cspC shows a positive autoregulation, which may occur via stabilization of its own mRNA.

Identificação de fatores de transcrição e sinais celulares que regulam a expressão do gene cspC em Caulobacter crescentus.