Goto-Seki A, Shirokane M, Masuda S, Tanaka K, Takahashi H
Institute of Molecular and Cellular Biosciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-0032, Japan.
Mol Microbiol. 1999 Nov;34(3):473-84. doi: 10.1046/j.1365-2958.1999.01608.x.
The chromosome of the cyanobacterium Synechococcus sp. PCC7942 contains at least one group 1 (rpoD1) and three group 2 (rpoD2, rpoD3 and rpoD4) sigma factor genes. In this study, we have analysed the structure of rpoD3 and rpoD4 and have shown that these genes are dispensable for growth at normal physiological conditions. An RNA polymerase core enzyme of the cyanobacterial strain was purified, reconstituted with the recombinant sigma factors (the rpoD1, rpoD3 and rpoD4 gene products), and the resultant holoenzymes were examined in vitro for transcription specificity. All of the holoenzymes recognized canonical promoters of Escherichia coli as well as cyanobacterial rrnA, cpcB1A1 P1a and rpoD1 promoters, although the three holoenzymes had some preference for specific promoters. These results suggest that group 1 as well as group 2 sigma factors of cyanobacteria may direct transcription initiation from the eubacterial consensus-type promoters containing the Pribnow -10 element, and we postulate that specificity crosstalk is a common characteristic among eubacterial group 1 and group 2 sigma factors. Phylogenetic analyses revealed that most group 2 sigma factors were positioned in one of four distinct clusters. The implication of the phylogenetic tree is also discussed in this paper.
蓝藻聚球藻属(Synechococcus sp.)PCC7942的染色体包含至少一个1类(rpoD1)和三个2类(rpoD2、rpoD3和rpoD4)的σ因子基因。在本研究中,我们分析了rpoD3和rpoD4的结构,并表明这些基因在正常生理条件下对生长并非必需。纯化了蓝藻菌株的RNA聚合酶核心酶,用重组σ因子(rpoD1、rpoD3和rpoD4基因产物)进行重组,然后在体外检测所得全酶的转录特异性。尽管这三种全酶对特定启动子有一定偏好,但所有全酶都能识别大肠杆菌的典型启动子以及蓝藻的rrnA、cpcB1A1 P1a和rpoD1启动子。这些结果表明,蓝藻的1类和2类σ因子可能指导从含有Pribnow -10元件的真细菌共有型启动子起始转录,并且我们推测特异性串扰是真细菌1类和2类σ因子的共同特征。系统发育分析表明,大多数2类σ因子位于四个不同簇中的一个。本文还讨论了系统发育树的含义。