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含主要σ因子的RNA聚合酶对蓝藻psbA启动子的特异性识别。

Specific recognition of the cyanobacterial psbA promoter by RNA polymerases containing principal sigma factors.

作者信息

Shibato J, Asayama M, Shirai M

机构信息

Laboratory of Molecular Microbiology, School of Agriculture, Ibaraki University, Ami, Inashiki, Ibaraki 300-0332, Japan.

出版信息

Biochim Biophys Acta. 1998 Nov 8;1442(2-3):296-303. doi: 10.1016/s0167-4781(98)00149-3.

Abstract

The psbA2 gene of a unicellular cyanobacterium, Microcystis aeruginosa K-81, encodes a D1 protein homolog in the reaction center of photosynthetic Photosystem II. To clarify the promoter recognition by a sigma factor of RNA polymerase, in vivo and in vitro analyses were performed for the photosynthetic gene. Although the specific transcript from the psbA2 promoter, whose sequence is of Escherichia coli consensus type, was observed in both cyanobacterium K-81 and E. coli cells, the expression was light-dependent in K-81 whereas it was constitutive in E. coli under the conditions of light and darkness (L/D). The specific psbA2-dependent transcripts were also detected in vitro by RNA polymerases containing the principal sigma factors, E. coli sigma70 and K-81 sigmaA1 (constitutively exists in K-81 grown under L/D cycles). Furthermore, a series of promoter fragments were constructed to confirm minimal cis elements for the in vitro psbA2 transcription. A -80 to +6 or -38 to +46 region, the sequences of which consisted of a core promoter (-38 to +6), was identified as the potential minimal cis element using the RNA polymerase fraction (*EsigmaA1) containing sigmaA1 partially purified from K-81. These results suggest that the psbA2 transcription with the minimal sequence was induced by the RNA polymerase (EsigmaA1) containing the principal sigma factor, sigmaA1, under both light and dark conditions in K-81.

摘要

单细胞蓝藻铜绿微囊藻K - 81的psbA2基因在光合光系统II的反应中心编码一种D1蛋白同源物。为了阐明RNA聚合酶的σ因子对启动子的识别,对该光合基因进行了体内和体外分析。尽管在蓝藻K - 81和大肠杆菌细胞中均观察到来自psbA2启动子的特异性转录本,其序列为大肠杆菌共有类型,但在K - 81中该表达是光依赖性的,而在大肠杆菌中,在光照和黑暗(L/D)条件下其表达是组成型的。含有主要σ因子大肠杆菌σ70和K - 81 σA1(在L/D循环下生长的K - 81中组成性存在)的RNA聚合酶在体外也检测到了特异性的psbA2依赖性转录本。此外,构建了一系列启动子片段以确认体外psbA2转录的最小顺式元件。使用从K - 81中部分纯化的含有σA1的RNA聚合酶组分(*EsigmaA1),将一个- 80至+6或- 38至+46区域(其序列由核心启动子(- 38至+6)组成)鉴定为潜在的最小顺式元件。这些结果表明,在K - 81中,无论光照还是黑暗条件下,含有主要σ因子σA1的RNA聚合酶(EsigmaA1)均可诱导具有最小序列psbA2的转录。

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