Wang Tao, Shen Gaozhong, Balasubramanian Ramakrishnan, McIntosh Lee, Bryant Donald A, Golbeck John H
Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802, USA.
J Bacteriol. 2004 Feb;186(4):956-67. doi: 10.1128/JB.186.4.956-967.2004.
The suf operon is composed of four genes (sufB, sufC, sufD, and sufS) and is highly conserved in the genomes of cyanobacteria. Open reading frame sll0088 in Synechocystis sp. strain PCC 6803 is located near the 5' end of the suf operon but is transcribed in the direction opposite that of the suf operon. We previously reported the isolation of two independent suppressor strains of C14S(PsaC) that mapped to sll0088 and restored photoautotrophic growth. The protein encoded by sll0088 has two significant features: (i) a DNA-binding domain near the N terminus and (ii) four highly conserved cysteine residues near the C terminus. The protein has high sequence similarity to transcription regulatory proteins with a conserved DNA-binding domain and can be classified in the DeoR family of helix-loop-helix proteins. The protein falls into a further subclass that contains a C-X(12)-C-X(13)-C-X(14)-C motif near the C terminus, which may represent a metal-binding site. The expressed Sll0088 protein harbored an iron-sulfur cluster as shown by optical and electron paramagnetic resonance spectroscopy. Compared to the wild type, expression levels of the sufBCDS genes were elevated when cells were grown under conditions of oxidative and iron stress and were even higher in a null mutant of Synechococcus sp. strain PCC 7002 in which the sll0088 homolog was insertionally inactivated. In agreement with the proposed role of the sufBCDS genes in iron metabolism, the growth rate of the null mutant was significantly higher than that of the wild type under iron-limiting conditions. We propose that the protein encoded by sll0088 is a transcriptional repressor of the suf operon, and we name the gene sufR.
suf操纵子由四个基因(sufB、sufC、sufD和sufS)组成,在蓝藻基因组中高度保守。集胞藻属PCC 6803菌株中的开放阅读框sll0088位于suf操纵子5'端附近,但转录方向与suf操纵子相反。我们之前报道了C14S(PsaC)的两个独立抑制菌株的分离,这些菌株定位于sll0088并恢复了光合自养生长。sll0088编码的蛋白质有两个显著特征:(i)N端附近有一个DNA结合结构域;(ii)C端附近有四个高度保守的半胱氨酸残基。该蛋白质与具有保守DNA结合结构域的转录调节蛋白具有高度序列相似性,可归类于螺旋-环-螺旋蛋白的DeoR家族。该蛋白质进一步归入一个亚类,在C端附近含有一个C-X(12)-C-X(13)-C-X(14)-C基序,这可能代表一个金属结合位点。如通过光学和电子顺磁共振光谱所示,表达的Sll0088蛋白含有一个铁硫簇。与野生型相比,当细胞在氧化和铁胁迫条件下生长时,sufBCDS基因的表达水平升高,在集球藻属PCC 7002菌株的一个无效突变体中甚至更高,其中sll0088同源物被插入失活。与sufBCDS基因在铁代谢中的推测作用一致,在铁限制条件下,无效突变体的生长速率显著高于野生型。我们提出sll0088编码的蛋白质是suf操纵子的转录抑制因子,并将该基因命名为sufR。