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转录延伸因子TFIIS参与拟南芥种子休眠。

Transcript elongation factor TFIIS is involved in arabidopsis seed dormancy.

作者信息

Grasser Marion, Kane Caroline M, Merkle Thomas, Melzer Michael, Emmersen Jeppe, Grasser Klaus D

机构信息

Department of Life Sciences, Aalborg University, Sohngaardsholmsvej 49, DK-9000 Aalborg, Denmark.

出版信息

J Mol Biol. 2009 Feb 27;386(3):598-611. doi: 10.1016/j.jmb.2008.12.066. Epub 2009 Jan 3.

Abstract

Transcript elongation factor TFIIS promotes efficient transcription by RNA polymerase II, since it assists in bypassing blocks during mRNA synthesis. While yeast cells lacking TFIIS are viable, inactivation of mouse TFIIS causes embryonic lethality. Here, we have identified a protein encoded in the Arabidopsis genome that displays a marked sequence similarity to TFIIS of other organisms, primarily within domains II and III in the C-terminal part of the protein. TFIIS is widely expressed in Arabidopsis, and a green fluorescent protein-TFIIS fusion protein localises specifically to the cell nucleus. When expressed in yeast cells lacking the endogenous TFIIS, Arabidopsis TFIIS partially complements the sensitivity of mutant cells to the nucleotide analog 6-azauridine, which is a typical characteristic of transcript elongation factors. We have characterised Arabidopsis lines harbouring T-DNA insertions in the coding sequence of TFIIS. Plants homozygous for T-DNA insertions are viable, and genomewide transcript profiling revealed that compared to control plants, a relatively small number of genes are differentially expressed in mutant plants. TFIIS(-/-) plants display essentially normal development, but they flower slightly earlier than control plants and show clearly reduced seed dormancy. Plants with RNAi-mediated knockdown of TFIIS expression also are affected in seed dormancy. Therefore, TFIIS plays a critical role in Arabidopsis seed development.

摘要

转录延伸因子TFIIS可促进RNA聚合酶II的高效转录,因为它有助于在mRNA合成过程中绕过转录障碍。虽然缺乏TFIIS的酵母细胞仍可存活,但小鼠TFIIS的失活会导致胚胎致死。在此,我们在拟南芥基因组中鉴定出一种编码蛋白,该蛋白与其他生物的TFIIS具有显著的序列相似性,主要在蛋白质C端的结构域II和III内。TFIIS在拟南芥中广泛表达,绿色荧光蛋白-TFIIS融合蛋白特异性定位于细胞核。当在缺乏内源性TFIIS的酵母细胞中表达时,拟南芥TFIIS可部分弥补突变细胞对核苷酸类似物6-氮尿苷的敏感性,这是转录延伸因子的典型特征。我们对在TFIIS编码序列中含有T-DNA插入的拟南芥株系进行了表征。T-DNA插入纯合的植株是可存活的,全基因组转录谱分析表明,与对照植株相比,突变植株中差异表达的基因数量相对较少。TFIIS(-/-)植株的发育基本正常,但它们比对照植株开花略早,且种子休眠明显降低。RNAi介导的TFIIS表达敲低的植株在种子休眠方面也受到影响。因此,TFIIS在拟南芥种子发育中起关键作用。

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