Dipartimento di Biotecnologie, Università di Verona, 15, Strada Le Grazie, I-37134 Verona, Italy.
Plant J. 2013 Mar;73(5):850-61. doi: 10.1111/tpj.12077. Epub 2012 Dec 28.
as1, for antenna size mutant 1, was obtained by insertion mutagenesis of the unicellular green alga Chlamydomonas reinhardtii. This strain has a low chlorophyll content, 8% with respect to the wild type, and displays a general reduction in thylakoid polypeptides. The mutant was found to carry an insertion into a homologous gene, prokaryotic arsenite transporter (ARSA), whose yeast and mammal counterparts were found to be involved in the targeting of tail-anchored (TA) proteins to cytosol-exposed membranes, essential for several cellular functions. Here we present the characterization in a photosynthetic organism of an insertion mutant in an ARSA-homolog gene. The ARSA1 protein was found to be localized in the cytosol, and yet its absence in as1 leads to a small chloroplast and a strongly decreased chlorophyll content per cell. ARSA1 appears to be required for optimal biogenesis of photosynthetic complexes because of its involvement in the accumulation of TOC34, an essential component of the outer chloroplast membrane translocon (TOC) complex, which, in turn, catalyzes the import of nucleus-encoded precursor polypeptides into the chloroplast. Remarkably, the effect of the mutation appears to be restricted to biogenesis of chlorophyll-binding polypeptides and is not compensated by the other ARSA homolog encoded by the C. reinhardtii genome, implying a non-redundant function.
as1 是通过插入诱变单细胞绿藻莱茵衣藻获得的。该菌株的叶绿素含量低,相对于野生型为 8%,并且类囊体多肽普遍减少。该突变体携带一个插入到同源基因 prokaryotic arsenite transporter (ARSA) 中,酵母和哺乳动物的对应物被发现参与将尾锚定 (TA) 蛋白靶向细胞质暴露的膜,这对几种细胞功能至关重要。在这里,我们在光合生物中介绍了 ARSA 同源基因插入突变体的特征。发现 ARSA1 蛋白定位于细胞质中,然而 as1 中其缺失导致叶绿体较小且每个细胞的叶绿素含量明显降低。ARSA1 似乎是光合作用复合物最佳生物发生所必需的,因为它参与了 TOC34 的积累,TOC34 是外叶绿体膜转运蛋白 (TOC) 复合物的必需组成部分,而 TOC 复合物又催化核编码前体多肽进入叶绿体。值得注意的是,该突变的影响似乎仅限于叶绿素结合多肽的生物发生,而不是由 C. reinhardtii 基因组编码的其他 ARSA 同源物补偿,这意味着其具有非冗余功能。