Sobczyk A, Schyns G, Tandeau de Marsac N, Houmard J
Départment de Biochimie et Génétique Moléculaire, Institut Pasteur, Paris, France.
EMBO J. 1993 Mar;12(3):997-1004. doi: 10.1002/j.1460-2075.1993.tb05740.x.
The cyanobacterium Calothrix sp. PCC 7601 can adapt its pigment content in response to changes in the incident light wavelength. It synthesizes, as major light-harvesting pigments, either phycocyanin 2 (PC2, encoded by the cpc2 operon) under red light or phycoerythrin (PE, encoded by the cpeBA operon) under green light conditions. The last step of the signal transduction pathway is characterized by a transcriptional control of the expression of these operons. Partially purified protein extracts were used in gel retardation assays and DNase I footprinting experiments to identify the factors that interact with the promoter region of the cpeBA operon. We found that two proteins, RcaA and RcaB, only detected in extracts of cells grown under green light, behave as positive transcriptional factors for the expression of the cpeBA operon. Treatment of the fractions containing RcaA and RcaB with alkaline phosphatase prevents the binding of RcaA but not of RcaB to the cpeBA promoter region. A post-translational modification of RcaA thus modulates its affinity for DNA.
蓝藻丝状绿球藻PCC 7601能够根据入射光波长的变化调整其色素含量。在红光条件下,它合成藻蓝蛋白2(PC2,由cpc2操纵子编码)作为主要的捕光色素;在绿光条件下,则合成藻红蛋白(PE,由cpeBA操纵子编码)。信号转导途径的最后一步以对这些操纵子表达的转录控制为特征。使用部分纯化的蛋白质提取物进行凝胶阻滞试验和DNase I足迹实验,以鉴定与cpeBA操纵子启动子区域相互作用的因子。我们发现,只有在绿光下生长的细胞提取物中检测到的两种蛋白质RcaA和RcaB,作为cpeBA操纵子表达的正转录因子。用碱性磷酸酶处理含有RcaA和RcaB的组分可阻止RcaA与cpeBA启动子区域的结合,但不能阻止RcaB的结合。因此,RcaA的翻译后修饰调节了其对DNA的亲和力。