National Key Laboratory of Plant Molecular Genetics, CAS Centre for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, 200032, China.
School of Life Sciences, Division of Life Sciences and Medicine, and Division of Molecular & Cell Biophysics, Hefei National Science Centre for Physical Sciences at the Microscale, University of Science and Technology of China, Hefei, 230027, China.
Plant Physiol. 2021 Aug 3;186(4):2003-2020. doi: 10.1093/plphys/kiab199.
The quality control system for messenger RNA (mRNA) is fundamental for cellular activities in eukaryotes. To elucidate the molecular mechanism of 3'-phosphoinositide-dependent protein kinase1 (PDK1), a master regulator that is essential throughout eukaryotic growth and development, we employed a forward genetic approach to screen for suppressors of the loss-of-function T-DNA insertion double mutant pdk1.1 pdk1.2 in Arabidopsis thaliana. Notably, the severe growth attenuation of pdk1.1 pdk1.2 was rescued by sop21 (suppressor of pdk1.1 pdk1.2), which harbors a loss-of-function mutation in PELOTA1 (PEL1). PEL1 is a homolog of mammalian PELOTA and yeast (Saccharomyces cerevisiae) DOM34p, which each form a heterodimeric complex with the GTPase HBS1 (HSP70 SUBFAMILY B SUPPRESSOR1, also called SUPERKILLER PROTEIN7, SKI7), a protein that is responsible for ribosomal rescue and thereby assures the quality and fidelity of mRNA molecules during translation. Genetic analysis further revealed that a dysfunctional PEL1-HBS1 complex failed to degrade the T-DNA-disrupted PDK1 transcripts, which were truncated but functional, and thus rescued the growth and developmental defects of pdk1.1 pdk1.2. Our studies demonstrated the functionality of a homologous PELOTA-HBS1 complex and identified its essential regulatory role in plants, providing insights into the mechanism of mRNA quality control.
信使 RNA(mRNA)的质量控制系统是真核细胞细胞活动的基础。为了阐明 3'-磷酸肌醇依赖性蛋白激酶 1(PDK1)的分子机制,PDK1 是真核生物生长和发育所必需的主要调节因子,我们采用正向遗传学方法筛选拟南芥 pdk1.1 pdk1.2 功能丧失 T-DNA 插入双突变体的抑制子。值得注意的是,pdk1.1 pdk1.2 的严重生长衰减被 sop21(pdk1.1 pdk1.2 的抑制子)挽救,sop21 携带 PELOTA1(PEL1)的功能丧失突变。PEL1 是哺乳动物 PELOTA 和酵母(酿酒酵母)DOM34p 的同源物,它们各自与 GTPase HBS1(HSP70 亚家族 B 抑制物 1,也称为超级杀手蛋白 7,SKI7)形成异二聚体复合物,该蛋白负责核糖体拯救,从而确保在翻译过程中 mRNA 分子的质量和保真度。遗传分析进一步表明,功能失调的 PEL1-HBS1 复合物不能降解 T-DNA 破坏的 PDK1 转录本,这些转录本虽然被截断但具有功能,从而挽救了 pdk1.1 pdk1.2 的生长和发育缺陷。我们的研究证明了同源 PELOTA-HBS1 复合物的功能,并确定了其在植物中的重要调节作用,为 mRNA 质量控制的机制提供了新的见解。