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拟南芥中的质体RNA聚合酶σ因子

Plastidic RNA polymerase sigma factors in Arabidopsis.

作者信息

Kanamaru K, Fujiwara M, Seki M, Katagiri T, Nakamura M, Mochizuki N, Nagatani A, Shinozaki K, Tanaka K, Takahashi H

机构信息

Laboratory of Molecular Genetics, University of Tokyo, Japan.

出版信息

Plant Cell Physiol. 1999 Aug;40(8):832-42. doi: 10.1093/oxfordjournals.pcp.a029612.

Abstract

In plant cells, plastid DNA is transcribed by at least two types of RNA polymerase, plastid-encoded RNA polymerase (PEP) and nuclear-encoded RNA polymerase (NEP). PEP is homologous to eubacterial transcription machinery, but its regulatory subunit, sigma (sigma) factor, is not encoded on the plastid DNA. We previously cloned the three nuclear-encoded sigma factor genes from Arabidopsis thaliana and designated them as sigA, sigB, and sigC. By means of RFLP mapping, sigA and sigB were mapped on chromosome I and sigC on the chromosome III. Based on comparison of the genomic structure of the three sig genes, intron sites in the 3' half of the genes were shown to be identical between sigB and sigC but divergent in sigA, consistent with the phylogenetic relevance of the three gene products. A transient expression assay of GFP fusions in Arabidopsis protoplasts showed that the N-termini of all three sig gene products functioned as chloroplast-targeting signals. We also constructed transgenic Arabidopsis lines harboring the sigA-promoter or the sigB-promoter uidA fusion. Both the sigA- and sigB-promoters were similarly activated at cotyledons, hypocotyls, rosette leaves, cauline leaves, sepals, and siliques but not at roots, seeds, or other flower organs. In addition, the two promoters were repeatedly activated in young seedlings under continuous light, possibly in an oscillated fashion.

摘要

在植物细胞中,质体DNA由至少两种类型的RNA聚合酶转录,即质体编码的RNA聚合酶(PEP)和核编码的RNA聚合酶(NEP)。PEP与真细菌转录机制同源,但其调节亚基,即σ(sigma)因子,并非由质体DNA编码。我们之前从拟南芥中克隆了三个核编码的σ因子基因,并将它们命名为sigA、sigB和sigC。通过限制性片段长度多态性(RFLP)作图,sigA和sigB定位于第一条染色体上,sigC定位于第三条染色体上。基于对这三个sig基因基因组结构的比较,发现sigB和sigC基因3'端的内含子位点相同,但与sigA不同,这与这三个基因产物的系统发育相关性一致。在拟南芥原生质体中对绿色荧光蛋白(GFP)融合蛋白进行的瞬时表达分析表明,所有三个sig基因产物的N端均作为叶绿体靶向信号发挥作用。我们还构建了携带sigA启动子或sigB启动子与uidA融合基因的转基因拟南芥品系。sigA和sigB启动子在子叶、下胚轴、莲座叶、茎生叶、萼片和角果中均有类似的激活,但在根、种子或其他花器官中没有。此外,在持续光照下,这两个启动子在幼苗中反复被激活,可能是以振荡的方式。

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