Kumakura Naoyoshi, Otsuki Hiroka, Tsuzuki Masayuki, Takeda Atsushi, Watanabe Yuichiro
Department of Life Sciences, Graduate School of Arts and Sciences, the University of Tokyo, Tokyo, Japan.
PLoS One. 2013 Nov 7;8(11):e79219. doi: 10.1371/journal.pone.0079219. eCollection 2013.
The RNA exosome is a multi-subunit complex that is responsible for 3' to 5' degradation and processing of cellular RNA. Rrp44/Dis3 is the catalytic center of the exosome in yeast and humans. However, the role of Rrp44/Dis3 homologs in plants is still unidentified. Here, we show that Arabidopsis AtRRP44A is the functional homolog of Rrp44/Dis3, is essential for plant viability and is required for RNA processing and degradation. We characterized AtRRP44A and AtRRP44B/SOV, two predicted Arabidopsis Rrp44/Dis3 homologs. AtRRP44A could functionally replace S. cerevisiae Rrp44/Dis3, but AtRRP44B/SOV could not. rrp44a knock-down mutants showed typical phenotypes of exosome function deficiency, 5.8S rRNA 3' extension and rRNA maturation by-product over-accumulation, but rrp44b mutants did not. Conversely, AtRRP44B/SOV mutants showed elevated levels of a selected mRNA, on which rrp44a did not have detectable effects. Although T-DNA insertion mutants of AtRRP44B/SOV had no obvious phenotype, those of AtRRP44A showed defects in female gametophyte development and early embryogenesis. These results indicate that AtRRP44A and AtRRP44B/SOV have independent roles for RNA turnover in plants.
RNA外切体是一种多亚基复合物,负责细胞RNA的3'至5'降解和加工。Rrp44/Dis3是酵母和人类中外切体的催化中心。然而,Rrp44/Dis3同源物在植物中的作用仍未明确。在此,我们表明拟南芥AtRRP44A是Rrp44/Dis3的功能同源物,对植物生存力至关重要,并且是RNA加工和降解所必需的。我们对两个预测的拟南芥Rrp44/Dis3同源物AtRRP44A和AtRRP44B/SOV进行了表征。AtRRP44A可以在功能上替代酿酒酵母的Rrp44/Dis3,但AtRRP44B/SOV不能。rrp44a敲低突变体表现出典型的外切体功能缺陷表型、5.8S rRNA 3'端延伸以及rRNA成熟副产物过度积累,但rrp44b突变体没有。相反,AtRRP44B/SOV突变体中一种选定的mRNA水平升高,而rrp44a对其没有可检测到的影响。虽然AtRRP44B/SOV的T-DNA插入突变体没有明显表型,但AtRRP44A的T-DNA插入突变体在雌配子体发育和早期胚胎发生中表现出缺陷。这些结果表明AtRRP44A和AtRRP44B/SOV在植物RNA周转中具有独立作用。