衣藻叶绿体中拟核分离与基因组完整性之间的动态相互作用

Dynamic Interplay between Nucleoid Segregation and Genome Integrity in Chlamydomonas Chloroplasts.

作者信息

Odahara Masaki, Kobayashi Yusuke, Shikanai Toshiharu, Nishimura Yoshiki

机构信息

Department of Botany, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan (M.O., Y.K., T.S., Y.N.); and.

Department of Life Science, College of Science, Rikkyo (St. Paul's) University, Toshima-ku, Tokyo 171-8501, Japan (M.O.).

出版信息

Plant Physiol. 2016 Dec;172(4):2337-2346. doi: 10.1104/pp.16.01533. Epub 2016 Oct 17.

Abstract

The chloroplast (cp) genome is organized as nucleoids that are dispersed throughout the cp stroma. Previously, a cp homolog of bacterial recombinase RecA (cpRECA) was shown to be involved in the maintenance of cp genome integrity by repairing damaged chloroplast DNA and by suppressing aberrant recombination between short dispersed repeats in the moss Physcomitrella patens Here, overexpression and knockdown analysis of cpRECA in the green alga Chlamydomonas reinhardtii revealed that cpRECA was involved in cp nucleoid dynamics as well as having a role in maintaining cp genome integrity. Overexpression of cpRECA tagged with yellow fluorescent protein or hemagglutinin resulted in the formation of giant filamentous structures that colocalized exclusively to chloroplast DNA and cpRECA localized to cp nucleoids in a heterogenous manner. Knockdown of cpRECA led to a significant reduction in cp nucleoid number that was accompanied by nucleoid enlargement. This phenotype resembled those of gyrase inhibitor-treated cells and monokaryotic chloroplast mutant cells and suggested that cpRECA was involved in organizing cp nucleoid dynamics. The cp genome also was destabilized by induced recombination between short dispersed repeats in cpRECA-knockdown cells and gyrase inhibitor-treated cells. Taken together, these results suggest that cpRECA and gyrase are both involved in nucleoid dynamics and the maintenance of genome integrity and that the mechanisms underlying these processes may be intimately related in C. reinhardtii cps.

摘要

叶绿体(cp)基因组以类核的形式组织,这些类核分散在整个cp基质中。此前,已证明细菌重组酶RecA的cp同源物(cpRECA)通过修复受损的叶绿体DNA以及抑制苔藓小立碗藓中短分散重复序列之间的异常重组,参与维持cp基因组的完整性。在此,对莱茵衣藻中cpRECA的过表达和敲低分析表明,cpRECA不仅参与cp基因组完整性的维持,还参与cp类核动态变化。用黄色荧光蛋白或血凝素标记的cpRECA过表达导致形成巨大的丝状结构,这些结构仅与叶绿体DNA共定位,并且cpRECA以异质方式定位于cp类核。cpRECA的敲低导致cp类核数量显著减少,并伴有类核增大。这种表型类似于用促旋酶抑制剂处理的细胞和单核叶绿体突变体细胞的表型,表明cpRECA参与组织cp类核动态变化。在cpRECA敲低细胞和促旋酶抑制剂处理的细胞中,短分散重复序列之间的诱导重组也使cp基因组不稳定。综上所述,这些结果表明cpRECA和促旋酶都参与类核动态变化和基因组完整性的维持,并且在莱茵衣藻cp中这些过程的潜在机制可能密切相关。

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