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Nucleus-encoded plastid sigma factor SIG3 transcribes specifically the psbN gene in plastids

机译:核编码质体sigma因子SIG3专门转录质体中的psbN基因

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摘要

We have investigated the function of one of the six plastid sigma-like transcription factors, sigma 3 (SIG3), by analysing two different Arabidopsis T-DNA insertion lines having disrupted SIG3 genes. Hybridization of wild-type and sig3 plant RNA to a plastid specific microarray revealed a strong reduction of the plastid psbN mRNA. The microarray result has been confirmed by northern blot analysis. The SIG3-specific promoter region has been localized on the DNA by primer extension and mRNA capping experiments. Results suggest tight regulation of psbN gene expression by a SIG3-PEP holoenzyme. The psbN gene is localized on the opposite strand of the psbB operon, between the psbT and psbH genes, and the SIG3-dependent psbN transcription produces antisense RNA to the psbT–psbH intergenic region. We show that this antisense RNA is not limited to the intergenic region, i.e. it does not terminate at the end of the psbN gene but extends as antisense transcript to cover the whole psbT coding region. Thus, by specific transcription initiation at the psbN gene promoter, SIG3-PEP holoenzyme could also influence the expression of the psbB operon by producing psbT antisense RNA.
机译:我们已经通过分析两个不同的拟南芥T-DNA插入线破坏了SIG3基因,调查了六个质体sigma样转录因子之一sigma 3(SIG3)的功能。野生型和sig3植物RNA与质体特异性微阵列的杂交显示,质体psbN mRNA显着降低。该微阵列结果已通过northern印迹分析确认。通过引物延伸和mRNA加帽实验,SIG3特异性启动子区域已定位在DNA上。结果表明SIG3-PEP全酶对psbN基因表达的严格调节。 psbN基因位于psbT和psbH基因之间的psbB操纵子的相反链上,而SIG3依赖的psbN转录产生了psbT–psbH基因间区域的反义RNA。我们显示出该反义RNA不限于基因间区域,即它不终止于psbN基因的末端,而是作为反义转录物延伸以覆盖整个psbT编码区。因此,通过在psbN基因启动子处的特异性转录起始,SIG3-PEP全酶也可以通过产生 psb T反义RNA来影响psbB操纵子的表达。

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