Department of Microbiology and Immunology, School of Medicine, State University of New York at Buffalo, Buffalo, New York 14214, USA.
RNA. 2011 May;17(5):865-77. doi: 10.1261/rna.2446311. Epub 2011 Mar 30.
Gene expression in the mitochondria of the kinetoplastid parasite Trypanosoma brucei is regulated primarily post-transcriptionally at the stages of RNA processing, editing, and turnover. The mitochondrial RNA-binding complex 1 (MRB1) is a recently identified multiprotein complex containing components with distinct functions during different aspects of RNA metabolism, such as guide RNA (gRNA) and mRNA turnover, precursor transcript processing, and RNA editing. In this study we examined the function of the MRB1 protein, Tb927.5.3010, which we term MRB3010. We show that MRB3010 is essential for growth of both procyclic form and bloodstream form life-cycle stages of T. brucei. Down-regulation of MRB3010 by RNAi leads to a dramatic inhibition of RNA editing, yet its depletion does not impact total gRNA levels. Rather, it appears to affect the editing process at an early stage, as indicated by the accumulation of pre-edited and small partially edited RNAs. MRB3010 is present in large (>20S) complexes and exhibits both RNA-dependent and RNA-independent interactions with other MRB1 complex proteins. Comparison of proteins isolated with MRB3010 tagged at its endogenous locus to those reported from other MRB1 complex purifications strongly suggests the presence of an MRB1 "core" complex containing five to six proteins, including MRB3010. Together, these data further our understanding of the function and composition of the imprecisely defined MRB1 complex.
线粒体基因表达在锥虫寄生虫布氏锥虫中主要在 RNA 加工、编辑和周转的转录后阶段进行调节。线粒体 RNA 结合复合物 1(MRB1)是一个最近被鉴定的多蛋白复合物,包含在 RNA 代谢的不同方面具有不同功能的成分,如指导 RNA(gRNA)和 mRNA 周转、前体转录本加工和 RNA 编辑。在这项研究中,我们研究了 MRB1 蛋白 Tb927.5.3010 的功能,我们将其称为 MRB3010。我们表明,MRB3010 对于布氏锥虫的前鞭毛体和血液体生活史阶段的生长都是必不可少的。通过 RNAi 下调 MRB3010 会导致 RNA 编辑的显著抑制,但它的耗竭并不影响总 gRNA 水平。相反,它似乎在早期影响编辑过程,如预编辑和小部分编辑 RNA 的积累所表明的那样。MRB3010 存在于大 (>20S) 复合物中,并与其他 MRB1 复合物蛋白表现出 RNA 依赖性和 RNA 非依赖性相互作用。与从其他 MRB1 复合物纯化中报道的蛋白相比,用 MRB3010 标记其内源基因座分离的蛋白进行比较强烈表明存在一个包含五到六个蛋白的 MRB1“核心”复合物,包括 MRB3010。这些数据共同进一步了解了功能和组成不精确定义的 MRB1 复合物。