Torabi Salar, Umate Pavan, Manavski Nikolay, Plöchinger Magdalena, Kleinknecht Laura, Bogireddi Hanumakumar, Herrmann Reinhold G, Wanner Gerhard, Schröder Wolfgang P, Meurer Jörg
Biozentrum der Ludwig-Maximilians-Universität München, Department Biologie I, 82152 Planegg-Martinsried, Germany.
Plant Cell. 2014 Mar;26(3):1183-99. doi: 10.1105/tpc.113.120444. Epub 2014 Mar 11.
The chloroplast-encoded low molecular weight protein PsbN is annotated as a photosystem II (PSII) subunit. To elucidate the localization and function of PsbN, encoded on the opposite strand to the psbB gene cluster, we raised antibodies and inserted a resistance cassette into PsbN in both directions. Both homoplastomic tobacco (Nicotiana tabacum) mutants psbN-F and psbN-R show essentially the same PSII deficiencies. The mutants are extremely light sensitive and failed to recover from photoinhibition. Although synthesis of PSII proteins was not altered significantly, both mutants accumulated only ∼25% of PSII proteins compared with the wild type. Assembly of PSII precomplexes occurred at normal rates, but heterodimeric PSII reaction centers (RCs) and higher order PSII assemblies were not formed efficiently in the mutants. The psbN-R mutant was complemented by allotopic expression of the PsbN gene fused to the sequence of a chloroplast transit peptide in the nuclear genome. PsbN represents a bitopic trans-membrane peptide localized in stroma lamellae with its highly conserved C terminus exposed to the stroma. Significant amounts of PsbN were already present in dark-grown seedling. Our data prove that PsbN is not a constituent subunit of PSII but is required for repair from photoinhibition and efficient assembly of the PSII RC.
叶绿体编码的低分子量蛋白PsbN被注释为光系统II(PSII)的一个亚基。为了阐明位于与psbB基因簇相反链上编码的PsbN的定位和功能,我们制备了抗体,并在两个方向上向PsbN中插入了一个抗性盒。同质性烟草(Nicotiana tabacum)突变体psbN-F和psbN-R都表现出基本相同的PSII缺陷。这些突变体对光极度敏感,无法从光抑制中恢复。虽然PSII蛋白的合成没有显著改变,但与野生型相比,两个突变体积累的PSII蛋白仅约为25%。PSII预复合物的组装以正常速率发生,但在突变体中异二聚体PSII反应中心(RCs)和更高阶的PSII组装体没有有效形成。psbN-R突变体通过在核基因组中异位表达与叶绿体转运肽序列融合的PsbN基因得到互补。PsbN是一种双位跨膜肽,定位于基质类囊体,其高度保守的C末端暴露于基质。在黑暗生长的幼苗中已经存在大量的PsbN。我们的数据证明,PsbN不是PSII的组成亚基,但对于从光抑制中修复和PSII RC的有效组装是必需的。