McDonnell Margaret M, Burkhart Sarah E, Stoddard Jerrad M, Wright Zachary J, Strader Lucia C, Bartel Bonnie
Department of BioSciences, Rice University, Houston, Texas, United States of America.
Department of Biology, Washington University at St. Louis, St. Louis, Missouri, United States of America.
PLoS One. 2016 Jan 29;11(1):e0148335. doi: 10.1371/journal.pone.0148335. eCollection 2016.
Peroxisomes are single-membrane bound organelles that are essential for normal development in plants and animals. In mammals and yeast, the peroxin (PEX) proteins PEX3 and PEX19 facilitate the early steps of peroxisome membrane protein (PMP) insertion and pre-peroxisome budding from the endoplasmic reticulum. The PEX3 membrane protein acts as a docking site for PEX19, a cytosolic chaperone for PMPs that delivers PMPs to the endoplasmic reticulum or peroxisomal membrane. PEX19 is farnesylated in yeast and mammals, and we used immunoblotting with prenylation mutants to show that PEX19 also is fully farnesylated in wild-type Arabidopsis thaliana plants. We examined insertional alleles disrupting either of the two Arabidopsis PEX19 isoforms, PEX19A or PEX19B, and detected similar levels of PEX19 protein in the pex19a-1 mutant and wild type; however, PEX19 protein was nearly undetectable in the pex19b-1 mutant. Despite the reduction in PEX19 levels in pex19b-1, both pex19a-1 and pex19b-1 single mutants lacked notable peroxisomal β-oxidation defects and displayed normal levels and localization of peroxisomal matrix and membrane proteins. The pex19a-1 pex19b-1 double mutant was embryo lethal, indicating a redundantly encoded critical role for PEX19 during embryogenesis. Expressing YFP-tagged versions of either PEX19 isoform rescued this lethality, confirming that PEX19A and PEX19B act redundantly in Arabidopsis. We observed that pex19b-1 enhanced peroxisome-related defects of a subset of peroxin-defective mutants, supporting a role for PEX19 in peroxisome function. Together, our data indicate that Arabidopsis PEX19 promotes peroxisome function and is essential for viability.
过氧化物酶体是由单层膜包裹的细胞器,对动植物的正常发育至关重要。在哺乳动物和酵母中,过氧化物酶体生物合成蛋白(PEX)中的PEX3和PEX19蛋白促进过氧化物酶体膜蛋白(PMP)插入以及过氧化物酶体前体从内质网出芽的早期步骤。PEX3膜蛋白作为PEX19的停靠位点,PEX19是一种PMP的胞质伴侣蛋白,可将PMP递送至内质网或过氧化物酶体膜。PEX19在酵母和哺乳动物中被法尼基化,我们通过对异戊二烯化突变体进行免疫印迹分析,结果表明PEX19在野生型拟南芥植株中也完全被法尼基化。我们检测了破坏拟南芥两种PEX19亚型(PEX19A或PEX19B)之一的插入突变体等位基因,发现在pex19a - 1突变体和野生型中PEX19蛋白水平相似;然而,在pex19b - 1突变体中几乎检测不到PEX19蛋白。尽管pex19b - 1中PEX19水平降低,但pex19a - 1和pex19b - 1单突变体均未表现出明显的过氧化物酶体β - 氧化缺陷,且过氧化物酶体基质和膜蛋白水平及定位均正常。pex19a - 1 pex19b - 1双突变体胚胎致死,这表明PEX19在胚胎发生过程中发挥着冗余编码的关键作用。表达带有黄色荧光蛋白(YFP)标签的任一PEX19亚型可挽救这种致死性,证实PEX19A和PEX19B在拟南芥中发挥冗余作用。我们观察到pex19b - 1增强了一部分过氧化物酶体生物合成缺陷型突变体的过氧化物酶体相关缺陷症状,这支持了PEX19在过氧化物酶体功能中发挥作用。总之,我们的数据表明拟南芥PEX19促进过氧化物酶体功能,对植株存活至关重要。