High-resolution map of plastid encoded polymerase binding patterns demonstrates a major role of transcription in chloroplast gene expression

High-resolution map of plastid encoded polymerase binding patterns demonstrates a major role of transcription in chloroplast gene expression
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质体编码聚合酶结合模式的高分辨率图证明转录在叶绿体基因表达中的主要作用

DOI:
10.1101/2022.01.18.476797
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发表时间:
2022
期刊:
bioRxiv
影响因子:
--
通讯作者:
Wierzbicki Andrzej T.
Wierzbicki Andrzej T.
中科院分区:
--
文献类型:
--
作者:
Palomar V. Miguel;Jaksich Sarah;Fujii Sho;Kucinski Jan;Wierzbicki Andrzej T.

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质体含有自己的基因组,由两种类型的 RNA 聚合酶转录。其中一种酶是由质体基因组编码的细菌型多亚基聚合酶。质体编码的 RNA 聚合酶 (PEP) 是光合作用相关蛋白质编码基因的有效表达所必需的。尽管 PEP 很重要,但其 DNA 结合位置尚未在全基因组范围内进行高分辨率研究。我们建立了一种高度特异性的方法,使用质体染色质免疫沉淀测序 (ptChIP-seq) 检测 PEP 与叶绿体 DNA 结合的全基因组模式。我们发现,在成熟的拟南芥叶绿体中,PEP 具有复杂的 DNA 结合模式,优先与编码 rRNA、tRNA 和光合蛋白子集的基因结合。 Sigma 因子 SIG2 和 SIG6 强烈影响 PEP 与 tRNA 基因子集的结合,并对整个基因组其余部分的 PEP 结合产生较温和的影响。 PEP 结合通常富集在转录起始位点周围的基因启动子上。最后,PEP 与 DNA 结合的水平与 RNA 积累的水平相关,这证明了 PEP 对叶绿体基因表达的影响。所提供的数据可通过公开的 Plastid 基因组可视化工具 (Plavisto) 获得,网址为 https://plavisto.mcdb.lsa.umich.edu/。
Plastids contain their own genomes, which are transcribed by two types of RNA polymerases. One of those enzymes is a bacterial‐type, multi‐subunit polymerase encoded by the plastid genome. The plastid‐encoded RNA polymerase (PEP) is required for efficient expression of genes encoding proteins involved in photosynthesis. Despite the importance of PEP, its DNA binding locations have not been studied on the genome‐wide scale at high resolution. We established a highly specific approach to detect the genome‐wide pattern of PEP binding to chloroplast DNA using plastid chromatin immunoprecipitation–sequencing (ptChIP‐seq). We found that in matureArabidopsis thalianachloroplasts, PEP has a complex DNA binding pattern with preferential association at genes encoding rRNA, tRNA, and a subset of photosynthetic proteins. Sigma factors SIG2 and SIG6 strongly impact PEP binding to a subset of tRNA genes and have more moderate effects on PEP binding throughout the rest of the genome. PEP binding is commonly enriched on gene promoters, around transcription start sites. Finally, the levels of PEP binding to DNA are correlated with levels of RNA accumulation, which demonstrates the impact of PEP on chloroplast gene expression. Presented data are available through a publicly available Plastid Genome Visualization Tool (Plavisto) at https://plavisto.mcdb.lsa.umich.edu/.
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