Department of Biochemistry and Cell Biology, Rice University, 6100 South Main Street, Houston, TX 77005, USA.
Plant Mol Biol. 2011 Sep;77(1-2):1-15. doi: 10.1007/s11103-011-9782-0. Epub 2011 May 8.
Mutations in peroxisome biogenesis proteins (peroxins) can lead to developmental deficiencies in various eukaryotes. PEX14 and PEX13 are peroxins involved in docking cargo-receptor complexes at the peroxisomal membrane, thus aiding in the transport of the cargo into the peroxisomal matrix. Genetic screens have revealed numerous Arabidopsis thaliana peroxins acting in peroxisomal matrix protein import; the viable alleles isolated through these screens are generally partial loss-of-function alleles, whereas null mutations that disrupt delivery of matrix proteins to peroxisomes can confer embryonic lethality. In this study, we used forward and reverse genetics in Arabidopsis to isolate four pex14 alleles. We found that all four alleles conferred reduced PEX14 mRNA levels and displayed physiological and molecular defects suggesting reduced but not abolished peroxisomal matrix protein import. The least severe pex14 allele, pex14-3, accumulated low levels of a C-terminally truncated PEX14 product that retained partial function. Surprisingly, even the severe pex14-2 allele, which lacked detectable PEX14 mRNA and PEX14 protein, was viable, fertile, and displayed residual peroxisome matrix protein import. As pex14 plants matured, import improved. Together, our data indicate that PEX14 facilitates, but is not essential for peroxisomal matrix protein import in plants.
过氧化物酶体生物发生蛋白(过氧化物酶体)的突变可导致各种真核生物的发育缺陷。PEX14 和 PEX13 是参与将货物受体复合物停靠在过氧化物酶体膜上的过氧化物酶体,从而有助于将货物运输到过氧化物酶体基质中。遗传筛选已经揭示了许多参与过氧化物酶体基质蛋白输入的拟南芥过氧化物酶体;通过这些筛选分离的存活等位基因通常是部分功能丧失等位基因,而破坏基质蛋白递送到过氧化物酶体的缺失突变可导致胚胎致死。在这项研究中,我们使用拟南芥的正向和反向遗传学来分离四个 pex14 等位基因。我们发现,所有四个等位基因都导致 PEX14 mRNA 水平降低,并表现出生理和分子缺陷,表明过氧化物酶体基质蛋白输入减少但未完全被阻断。pex14-3 等位基因是最不严重的,它积累了低水平的 C 末端截断的 PEX14 产物,该产物保留了部分功能。令人惊讶的是,即使是严重的 pex14-2 等位基因,其缺乏可检测的 PEX14 mRNA 和 PEX14 蛋白,也是存活的、可育的,并表现出残留的过氧化物酶体基质蛋白输入。随着 pex14 植物的成熟,输入得到改善。总之,我们的数据表明 PEX14 促进但不是必需的植物过氧化物酶体基质蛋白输入。