The rpoD1 gene product is a principal sigma factor of RNA polymerase in Microcystis aeruginosa K-81.
The rpoD1 gene product is a principal sigma factor of RNA polymerase in Microcystis aeruginosa K-81.
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rpoD1 基因产物是铜绿微囊藻 K-81 中 RNA 聚合酶的主要 Sigma 因子。
DOI：
10.1093/oxfordjournals.jbchem.a021475
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发表时间：
1996
影响因子：
2.7
通讯作者：
M. Shirai
中科院分区：
文献类型：
--
作者：
M. Asayama;H. Suzuki;A. Sato;T. Aida;K. Tanaka;H. Takahashi;M. Shirai
我们对从铜绿微囊藻（Microcystis aeruginosa K-81）中分离的RpoD1基因编码的RpoD1蛋白进行了分子表征。推导出的RpoD1的氨基酸序列（416 aa, 48,871 Da）与真细菌RpoD家族（大肠杆菌sigma 70同源）的蛋白质具有广泛的相似性。我们从大肠杆菌中过量生产并纯化了RpoD1 （54 kDa）。生物学和生化分析表明，RpoD1具有与大肠杆菌sigma 70同源的功能：(i) RpoD1蛋白与大肠杆菌YN543的rpoD突变体（rpoD285）互补；（ii）由大肠杆菌核心酶重组的异源RNA聚合酶全酶和重组RpoD1特异性转录自大肠杆菌启动子。此外，用Western blot方法对聚珠球菌菌株PCC 7942 RpoD1 （sigma 70型的主sigma因子）的抗血清进行分析，结果表明M. aeruginosa K-81 RpoD1 （sigma A1）是影响指数细胞生长的sigma亚基的主sigma因子。
We performed molecular characterization of the RpoD1 protein encoded by the rpoD1 gene isolated from a cyanobacterium, Microcystis aeruginosa K-81. The deduced amino acid sequence (416 aa, 48,871 Da) of RpoD1 exhibited extensive similarity to those of proteins of the eubacterial RpoD family (Escherichia coli sigma 70 homologs). We overproduced and purified RpoD1 (54 kDa) from E. coli. Biological and biochemical analyses suggested that RpoD1 has a function homologous to that of E. coli sigma 70 as follows: (i) the RpoD1 protein complemented an rpoD mutant of E. coli strain YN543 (rpoD285) and (ii) the heterologous RNA polymerase holoenzyme reconstituted from the E. coli core enzyme and recombinant RpoD1 was specifically transcribed from E. coli promoters. Furthermore, Western blot analysis with antiserum against Synechococcus sp. strain PCC 7942 RpoD1 (a principal sigma factor of the sigma 70 type) indicated that M. aeruginosa K-81 RpoD1 (sigma A1) is the principal sigma factor, which is a major component of the sigma subunit on exponential cell growth.