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Evidence for redox regulation of the transcription factor NtcA acting both as an activator and a repressor in the cyanobacterium Anabaena PCC 7120.

机译:蓝藻鱼腥藻PCC 7120中氧化因子调节转录因子NtcA既作为激活剂又是抑制因子的证据。

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摘要

NtcA has been identified as a nitrogen-responsive regulatory protein required for nitrogen assimilation and heterocyst differentiation in cyanobacteria. It is proposed that NtcA functions through the formation of DNA-protein complexes with its specific target sequence within the promoter regions of the regulated genes. In vitro, NtcA of Anabaena PCC 7120 binds to upstream regions of the genes whose products are involved in nitrogen assimilation, but also to the upstream region of rbcLS (carbon-fixation gene), xisA (encoding a site-specific recombinase expressed during heterocyst differentiation) and ntcA (encoding NtcA itself). However, the mechanism by which NtcA serves as a critical regulator for such diverse processes is not understood. With the use of electrophoretic mobility shift assays, NtcA from Anabaena PCC 7120 was here shown to interact with the promoter sequence of the gor gene, encoding glutathione reductase, thereby providing a novel example of NtcA's acting as a repressor, previously found only for the rbcLS gene. Furthermore we demonstrate that the binding of DNA by NtcA is regulated in vitro by a redox-dependent mechanism involving cysteine residues of the NtcA protein. These findings suggest that NtcA is a transcriptional regulator that responds not only to the nitrogen status but also to the cellular redox status, a function that might be particularly significant during heterocyst differentiation.
机译:NtcA已被确定为蓝细菌中氮同化和异囊分化所需的氮响应调节蛋白。提出NtcA通过在调节基因的启动子区域内形成具有特定靶序列的DNA-蛋白质复合物来发挥功能。在体外,鱼腥藻PCC 7120的NtcA结合其产物参与氮同化的基因的上游区域,但也结合到rbcLS(碳固定基因),xisA(编码在异胞藻分化过程中表达的位点特异性重组酶)的上游区域。 )和ntcA(编码NtcA本身)。但是,尚不了解NtcA充当此类多样化过程的关键调节剂的机制。使用电泳迁移率迁移分析,来自Anabaena PCC 7120的NtcA在这里显示与gor基因的启动子序列相互作用,编码谷胱甘肽还原酶,从而提供了一个NtcA充当阻遏物的新例子,以前仅在rbcLS中发现基因。此外,我们证明了NtcA与DNA的结合在体外受氧化还原依赖性机制的调节,该机制涉及NtcA蛋白的半胱氨酸残基。这些发现表明,NtcA是一种转录调节因子,它不仅对氮状态而且还对细胞氧化还原状态有反应,这种功能在异型囊肿分化过程中可能特别重要。

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