Oikawa K, Tanaka K, Takahashi H
Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.
Gene. 1998 Apr 14;210(2):277-85. doi: 10.1016/s0378-1119(98)00075-4.
A nuclear gene, sigA, that encodes a sigma factor for chloroplast RNA polymerase has previously been identified and characterized in the primitive red alga Cyanidium caldarium strain RK-1. Southern hybridization analysis indicated the presence of two additional sigma factor genes, which have now been cloned and shown to encode virtually identical proteins that are homologous to eubacterial sigma factors. These genes, which are also present in the nuclear genome, have therefore been named sigB and sigC. The substantial sequence similarity of sigB and sigC to sigA of the same strain as well as to cyanobacterial principal sigma factors and other chloroplast sigma factors strongly suggests that the nuclear genome of C. caldarium contains three genes that encode two types of chloroplast sigma factors. Each of the three recombinant Sig proteins showed sigma factor activity in vitro when combined with the Escherichia coli RNA polymerase core enzyme. Northern blot analysis revealed that, whereas the overall abundance of sigA transcripts was not affected by light, the amount of sigB and sigC mRNAs was greater in the light than in the dark. Thus, multiple sigma factors appear to contribute to light-regulated gene expression in the chloroplast.
一个编码叶绿体RNA聚合酶σ因子的核基因sigA,先前已在原始红藻嗜热蓝细菌RK-1菌株中被鉴定和表征。Southern杂交分析表明存在另外两个σ因子基因,现已克隆出这两个基因,并显示它们编码与真细菌σ因子同源的几乎相同的蛋白质。这些同样存在于核基因组中的基因,因此被命名为sigB和sigC。sigB和sigC与同一菌株的sigA以及蓝细菌主要σ因子和其他叶绿体σ因子具有显著的序列相似性,这强烈表明嗜热蓝细菌的核基因组包含三个编码两种类型叶绿体σ因子的基因。当与大肠杆菌RNA聚合酶核心酶结合时,三种重组Sig蛋白在体外均表现出σ因子活性。Northern印迹分析显示,虽然sigA转录本的总体丰度不受光照影响,但sigB和sigC mRNA的量在光照下比在黑暗中更多。因此,多种σ因子似乎参与了叶绿体中光调节的基因表达。