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A nuclear-encoded protein mTERF6 mediates transcription termination of rpoA polycistron for plastid-encoded RNA polymerase-dependent chloroplast gene expression and chloroplast development

机译:核编码蛋白mTERF6介导rpoA多顺反子的转录终止用于质体编码的RNA聚合酶依赖性叶绿体基因表达和叶绿体发育

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摘要

The expression of plastid genes is regulated by two types of DNA-dependent RNA polymerases, plastid-encoded RNA polymerase (PEP) and nuclear-encoded RNA polymerase (NEP). The plastid rpoA polycistron encodes a series of essential chloroplast ribosome subunits and a core subunit of PEP. Despite the functional importance, little is known about the regulation of rpoA polycistron. In this work, we show that mTERF6 directly associates with a 3′-end sequence of rpoA polycistron in vitro and in vivo, and that absence of mTERF6 promotes read-through transcription at this site, indicating that mTERF6 acts as a factor required for termination of plastid genes’ transcription in vivo. In addition, the transcriptions of some essential ribosome subunits encoded by rpoA polycistron and PEP-dependent plastid genes are reduced in the mterf6 knockout mutant. RpoA, a PEP core subunit, accumulates to about 50% that of the wild type in the mutant, where early chloroplast development is impaired. Overall, our functional analyses of mTERF6 provide evidence that it is more likely a factor required for transcription termination of rpoA polycistron, which is essential for chloroplast gene expression and chloroplast development.
机译:质体基因的表达受两种类型的DNA依赖性RNA聚合酶调节,质体编码的RNA聚合酶(PEP)和核编码的RNA聚合酶(NEP)。质体rpoA多顺反子编码一系列必需的叶绿体核糖体亚基和PEP的核心亚基。尽管功能上很重要,但对rpoA多顺反子的调控知之甚少。在这项工作中,我们表明mTERF6在体外和体内与rpoA多顺反子的3'末端序列直接缔合,而mTERF6的缺失会促进该位点的通读转录,表明mTERF6充当终止所需的因子质体基因在体内的转录此外,在mterf6基因敲除突变体中,rpoA多顺反子和PEP依赖的质体基因编码的一些必需核糖体亚基的转录降低。 RpoA是PEP的核心亚基,在突变体中积累了约50%的野生型,而早期叶绿体发育受到了破坏。总体而言,我们对mTERF6的功能分析提供了证据,表明它很可能是rpoA多顺反子转录终止所需的因子,这对叶绿体基因表达和叶绿体发育至关重要。

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