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PAP 基因是叶绿体发育的组织和细胞特异性标记物。

PAP genes are tissue- and cell-specific markers of chloroplast development.

机构信息

LPCV, CEA, CNRS, INRA, Université Grenoble-Alpes, BIG, 38000, Grenoble, France.

出版信息

Planta. 2018 Sep;248(3):629-646. doi: 10.1007/s00425-018-2924-8. Epub 2018 May 31.

Abstract

Expression of PAP genes is strongly coordinated and represents a highly selective cell-specific marker associated with the development of chloroplasts in photosynthetically active organs of Arabidopsis seedlings and adult plants. Transcription in plastids of plants depends on the activity of phage-type single-subunit nuclear-encoded RNA polymerases (NEP) and a prokaryotic multi-subunit plastid-encoded RNA polymerase (PEP). PEP is comprised of the core subunits α, β, β' and β″ encoded by rpoA, rpoB/C/C genes located on the plastome. This core enzyme needs to interact with nuclear-encoded sigma factors for proper promoter recognition. In chloroplasts, the core enzyme is surrounded by additional 12 nuclear-encoded subunits, all of eukaryotic origin. These PEP-associated proteins (PAPs) were found to be essential for chloroplast biogenesis as Arabidopsis inactivation mutants for each of them revealed albino or pale-green phenotypes. In silico analysis of transcriptomic data suggests that PAP genes represent a tightly controlled regulon, whereas wetlab data are sparse and correspond to the expression of individual genes mostly studied at the seedling stage. Using RT-PCR, transient, and stable expression assays of PAP promoter-GUS-constructs, we do provide, in this study, a comprehensive expression catalogue for PAP genes throughout the life cycle of Arabidopsis. We demonstrate a selective impact of light on PAP gene expression and uncover a high tissue specificity that is coupled to developmental progression especially during the transition from skotomorphogenesis to photomorphogenesis. Our data imply that PAP gene expression precedes the formation of chloroplasts rendering PAP genes a tissue- and cell-specific marker of chloroplast biogenesis.

摘要

PAP 基因的表达受到严格调控,是一种与拟南芥幼苗和成年植物光合作用器官中叶绿体发育相关的高度选择性细胞特异性标记物。植物质体中的转录依赖于噬菌体型单亚基核编码 RNA 聚合酶 (NEP) 和原核多亚基质体编码 RNA 聚合酶 (PEP) 的活性。PEP 由 rpoA、rpoB/C/C 基因编码的核心亚基α、β、β'和β"组成,这些基因位于质体基因组上。这种核心酶需要与核编码的 σ 因子相互作用,以正确识别启动子。在叶绿体中,核心酶被额外的 12 个核编码亚基包围,这些亚基均具有真核生物的起源。研究发现,这些 PEP 相关蛋白 (PAP) 对于叶绿体的生物发生是必不可少的,因为拟南芥的每个失活突变体都表现出白化或淡绿色表型。基于转录组数据分析的计算分析表明,PAP 基因代表一个受到严格调控的调控单元,而湿实验室数据则很稀疏,且主要对应于在幼苗阶段研究的单个基因的表达。通过 RT-PCR、瞬时和稳定表达分析 PAP 启动子-GUS-构建体,我们在本研究中提供了拟南芥整个生命周期中 PAP 基因的全面表达目录。我们证明了光对 PAP 基因表达的选择性影响,并揭示了高度的组织特异性,这种特异性与发育进程相关,特别是在从暗形态建成到光形态建成的转变过程中。我们的数据表明,PAP 基因的表达先于叶绿体的形成,这使得 PAP 基因成为叶绿体生物发生的组织和细胞特异性标记物。

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