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S1 结构域包含的 STF 调节拟南芥中质体转录和叶绿体生物发生。

S1 domain-containing STF modulates plastid transcription and chloroplast biogenesis in Nicotiana benthamiana.

机构信息

Department of Systems Biology, Yonsei University, Seoul, Korea.

出版信息

New Phytol. 2012 Jan;193(2):349-63. doi: 10.1111/j.1469-8137.2011.03941.x. Epub 2011 Nov 4.

Abstract

• In this study, we examined the biochemical and physiological functions of Nicotiana benthamiana S1 domain-containing Transcription-Stimulating Factor (STF) using virus-induced gene silencing (VIGS), cosuppression, and overexpression strategies. • STF : green fluorescent protein (GFP) fusion protein colocalized with sulfite reductase (SiR), a chloroplast nucleoid-associated protein also present in the stroma. Full-length STF and its S1 domain preferentially bound to RNA, probably in a sequence-nonspecific manner. • STF silencing by VIGS or cosuppression resulted in severe leaf yellowing caused by disrupted chloroplast development. STF deficiency significantly perturbed plastid-encoded multimeric RNA polymerase (PEP)-dependent transcript accumulation. Chloroplast transcription run-on assays revealed that the transcription rate of PEP-dependent plastid genes was reduced in the STF-silenced leaves. Conversely, the exogenously added recombinant STF protein increased the transcription rate, suggesting a direct role of STF in plastid transcription. Etiolated seedlings of STF cosuppression lines showed defects in the light-triggered transition from etioplasts to chloroplasts, accompanied by reduced light-induced expression of plastid-encoded genes. • These results suggest that STF plays a critical role as an auxiliary factor of the PEP transcription complex in the regulation of plastid transcription and chloroplast biogenesis in higher plants.

摘要

• 在本研究中,我们利用病毒诱导的基因沉默(VIGS)、共抑制和过表达策略,研究了黄花烟 S1 结构域转录刺激因子(STF)的生化和生理功能。

• STF:绿色荧光蛋白(GFP)融合蛋白与亚硫酸还原酶(SiR)共定位,SiR 是一种质体核仁相关蛋白,也存在于基质中。全长 STF 及其 S1 结构域优先与 RNA 结合,可能以非序列特异性的方式结合。

• 通过 VIGS 或共抑制沉默 STF 会导致叶绿体发育受阻,叶片严重黄化。STF 缺失显著干扰质体编码的多聚 RNA 聚合酶(PEP)依赖性转录本积累。质体转录启动子分析显示,STF 沉默叶片中 PEP 依赖性质体基因的转录速率降低。相反,外源添加的重组 STF 蛋白增加了转录速率,表明 STF 直接参与质体转录。STF 共抑制系的黄化幼苗在由黄化质体向叶绿体的光触发转变过程中出现缺陷,同时质体编码基因的光诱导表达减少。

• 这些结果表明,STF 作为 PEP 转录复合物的辅助因子,在高等植物质体转录和叶绿体生物发生的调控中发挥着关键作用。

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