Institute for Protein Research, Osaka University, Suita, 565-0871, Japan.
Plant Cell. 2022 Oct 27;34(11):4569-4582. doi: 10.1093/plcell/koac240.
The complexes translocon at the outer envelope membrane of chloroplasts and translocon at the inner envelope membrane of chloroplasts (TIC) mediate preprotein translocation across the chloroplast outer and inner envelope membranes, respectively. Tic20, Tic56, Tic100, and Tic214 form a stable one-megadalton TIC whose function is essential for Arabidopsis thaliana and Chlamydomonas reinhardtii. Tic20 plays a central role in preprotein translocation by forming a protein-conducting channel. Tic56, Tic100, and Tic214 are also indispensable for TIC function, but whether other components are required for this process remain unclear. Here, we demonstrate that a 12-kDa protein named Tic12 is part of the TIC in A. thaliana and participates in preprotein translocation across the inner envelope membrane. Tic12 was tightly associated with the TIC but disassociated under high-salt conditions in combination with Triton X-100. Site-specific UV crosslinking experiments revealed that Tic12 and Tic20 directly interact with the transit peptide of a translocating preprotein. The tic12 null mutants are albino and seedling lethal, similar to the other tic null mutants. Tic12 and Tic20 were also involved in preprotein translocation in (Pisum sativum) pea chloroplasts. Thus, Tic12 is an essential constituent that forms the functional core together with Tic20 in the one-megadalton TIC.
叶绿体的外膜转位复合物和叶绿体的内膜转位复合物(TIC)分别介导前体蛋白穿过叶绿体的外膜和内膜的转移。Tic20、Tic56、Tic100 和 Tic214 形成一个稳定的 1 兆道尔顿 TIC,其功能对于拟南芥和莱茵衣藻是必不可少的。Tic20 通过形成蛋白质导通道在前体蛋白转运中发挥核心作用。Tic56、Tic100 和 Tic214 对于 TIC 功能也是不可或缺的,但该过程是否还需要其他成分尚不清楚。在这里,我们证明了一种名为 Tic12 的 12kDa 蛋白是拟南芥 TIC 的一部分,并参与了前体蛋白穿过内膜的转运。Tic12 与 TIC 紧密相关,但在高盐条件下与 Triton X-100 结合时会解离。特异性 UV 交联实验表明,Tic12 和 Tic20 与转运前体蛋白的易位肽直接相互作用。tic12 缺失突变体是白化体和幼苗致死的,与其他 tic 缺失突变体相似。Tic12 和 Tic20 也参与了豌豆叶绿体的前体蛋白转运。因此,Tic12 是一个必需的组成部分,与 Tic20 一起形成功能核心,存在于 1 兆道尔顿 TIC 中。