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质体缺陷蛋白 1 对于质体编码 RNA 聚合酶核心亚基 β 的积累和叶绿体发育是必需的。

Plastid Deficient 1 Is Essential for the Accumulation of Plastid-Encoded RNA Polymerase Core Subunit β and Chloroplast Development in .

机构信息

Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.

College of Life Science, University of Chinese Academy of Sciences, Beijing 100049, China.

出版信息

Int J Mol Sci. 2021 Dec 20;22(24):13648. doi: 10.3390/ijms222413648.

Abstract

Plastid-encoded RNA polymerase (PEP)-dependent transcription is an essential process for chloroplast development and plant growth. It is a complex event that is regulated by numerous nuclear-encoded proteins. In order to elucidate the complex regulation mechanism of PEP activity, identification and characterization of PEP activity regulation factors are needed. Here, we characterize Plastid Deficient 1 (PD1) as a novel regulator for PEP-dependent gene expression and chloroplast development in . The gene encodes a protein that is conserved in photoautotrophic organisms. The   mutant showed albino and seedling-lethal phenotypes. The plastid development in the mutant was arrested. The PD1 protein localized in the chloroplasts, and it colocalized with nucleoid protein TRXz. RT-quantitative real-time PCR, northern blot, and run-on analyses indicated that the PEP-dependent transcription in the mutant was dramatically impaired, whereas the nuclear-encoded RNA polymerase-dependent transcription was up-regulated. The yeast two-hybrid assays and coimmunoprecipitation experiments showed that the PD1 protein interacts with PEP core subunit β (PEP-β), which has been verified to be essential for chloroplast development. The immunoblot analysis indicated that the accumulation of PEP-β was barely detected in the mutant, whereas the accumulation of the other essential components of the PEP complex, such as core subunits α and β', were not affected in the mutant. These observations suggested that the PD1 protein is essential for the accumulation of PEP-β and chloroplast development in , potentially by direct interaction with PEP-β.

摘要

质体编码 RNA 聚合酶(PEP)依赖性转录是叶绿体发育和植物生长所必需的过程。它是一个复杂的事件,受许多核编码蛋白的调节。为了阐明 PEP 活性的复杂调节机制,需要鉴定和表征 PEP 活性调节因子。在这里,我们将 Plastid Deficient 1(PD1)鉴定为 中 PEP 依赖性基因表达和叶绿体发育的新型调节因子。基因编码的蛋白在自养生物中保守。突变体表现出白化和幼苗致死表型。突变体中的质体发育被阻断。PD1 蛋白定位于叶绿体中,与核蛋白 TRXz 共定位。RT-定量实时 PCR、 northern blot 和转录分析表明,突变体中的 PEP 依赖性转录显著受损,而核编码 RNA 聚合酶依赖性转录上调。酵母双杂交试验和共免疫沉淀实验表明,PD1 蛋白与 PEP 核心亚基 β(PEP-β)相互作用,PEP-β 已被证实对叶绿体发育至关重要。免疫印迹分析表明,在突变体中几乎检测不到 PEP-β 的积累,而 PEP 复合物的其他必需成分,如核心亚基 α 和 β',在突变体中没有受到影响。这些观察结果表明,PD1 蛋白对于 PEP-β 的积累和 的叶绿体发育是必需的,可能通过与 PEP-β 的直接相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6389/8705867/f8ebc95e3caf/ijms-22-13648-g001a.jpg

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