Redox control of ntcA gene expression in Synechocystis sp. PCC 6803. Nitrogen availability and electron transport regulate the levels of the NtcA protein.
In this work we have studied the influence of the cellular redox status in the expression of the Synechocystis sp. PCC 6803 ntcA gene. Two different ntcA transcripts with different 5' ends were detected, depending on the different dark/light or nitrogen availability conditions. Accumulation of a 0.8-kb ntcA message was light and nitrogen dependent, whereas a longer 1.2-kb ntcA transcript was neither light nor nitrogen regulated. NtcA protein levels increased concomitantly with the accumulation of the 0.8-kb ntcA transcript. The light-dependent accumulation of the ntcA gene and the NtcA protein was sensitive to electron transport inhibitors. In addition, Glc-grown Synechocystis sp. cells showed a similar ntcA expression pattern in darkness to that observed under illumination. These data suggested that electron transport, and not light per se may regulate ntcA gene expression. Primer extension analysis, together with gel mobility-shift assays, demonstrated that in vitro, the Synechocystis sp. NtcA protein specifically bound to the putative promoter region from the light/nitrogen-dependent ntcA transcript but not to that from the constitutive 1.2-kb ntcA mRNA. Band-shift experiments carried out in the presence of thiol oxidizing/modifiying agents and different reducing/oxidizing conditions suggested that NtcA binding to its own promoter was under a thiol-dependent redox mechanism. Our results suggest that the cellular redox status plays a central role in the autoregulatory mechanism of the NtcA protein.
Alfonso
M
M
Unité Mixte de Recherche, 8543 Photorégulation et Dynamique des Membranes Végetales, Centre National de la Recherche Scientifique, Ecole Normale Supériéure, 46 rue d'Ulm, 75230 Paris cedex 05, France. alfonso@ead.csic.es
Perewoska
I
I
Kirilovsky
D
D
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D002478:Cells, Cultured; D000458:Cyanobacteria / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003624:Darkness; D015964:Gene Expression Regulation, Bacterial; D008027:Light; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D014157:Transcription Factors; C075617:ntcA protein, Synechococcus