Department of Photochemistry and Molecular Science, Angström Laboratories, Uppsala University, Uppsala, Sweden.
Appl Environ Microbiol. 2010 Feb;76(3):880-90. doi: 10.1128/AEM.02521-09. Epub 2009 Dec 18.
The filamentous, heterocystous, nitrogen-fixing cyanobacterium Nostoc sp. strain PCC 7120 may contain, depending on growth conditions, up to two hydrogenases directly involved in hydrogen metabolism. HypC is one out of at least seven auxiliary gene products required for synthesis of a functional hydrogenase, specifically involved in the maturation of the large subunit. In this study we present a protein, CalA (Alr0946 in the genome), belonging to the transcription regulator family AbrB, which in protein-DNA assays was found to interact with the upstream region of hypC. Transcriptional investigations showed that calA is cotranscribed with the downstream gene alr0947, which encodes a putative protease from the abortive infection superfamily, Abi. CalA was shown to interact specifically not only with the upstream region of hypC but also with its own upstream region, acting as a repressor on hypC. The bidirectional hydrogenase activity was significantly downregulated when CalA was overexpressed, demonstrating a correlation with the transcription factor, either direct or indirect. In silico studies showed that homologues to both CalA and Alr0947 are highly conserved proteins within cyanobacteria with very similar physical organizations of the corresponding structural genes. Possible functions of the cotranscribed downstream protein Alr0947 are presented. In addition, we present a three-dimensional (3D) model of the DNA binding domain of CalA and putative DNA binding mechanisms are discussed.
丝状异形胞固氮蓝藻 Nostoc sp. 株 PCC 7120 可能根据生长条件包含多达两种直接参与氢代谢的氢化酶。HypC 是合成功能性氢化酶所需的至少七种辅助基因产物之一,特别是参与大亚基的成熟。在这项研究中,我们提出了一种属于转录调节因子家族 AbrB 的蛋白 CalA(基因组中的 Alr0946),在蛋白-DNA 分析中发现它与 hypC 的上游区域相互作用。转录研究表明,calA 与下游基因 alr0947 共转录,该基因编码来自 abortive infection 超家族的假定蛋白酶 Abi。CalA 不仅与 hypC 的上游区域特异性相互作用,而且与自身的上游区域相互作用,作为 hypC 的抑制剂。当 CalA 过表达时,双向氢化酶活性显著下调,表明与转录因子直接或间接相关。计算机研究表明,CalA 和 Alr0947 的同源物在蓝藻中是高度保守的蛋白质,其相应结构基因的物理组织非常相似。提出了共转录下游蛋白 Alr0947 的可能功能。此外,我们还展示了 CalA 的 DNA 结合域的三维(3D)模型,并讨论了假定的 DNA 结合机制。