Pollard V W, Harris M E, Hajduk S L
Department of Biochemistry, University of Alabama, School of Medicine, Birmingham 35294.
EMBO J. 1992 Dec;11(12):4429-38. doi: 10.1002/j.1460-2075.1992.tb05543.x.
The aim of this study was to identify multicomponent complexes involved in kinetoplastid mitochondrial mRNA editing. Mitochondrial extracts from Trypanosoma brucei were fractionated on 10-30% glycerol gradients and assayed for RNAs and activities potentially involved in editing, including pre-edited mRNA, guide RNA (gRNA), endonuclease, terminal uridylyltransferase (TUTase), RNA ligase and gRNA-mRNA chimera-forming activities. These experiments suggest that two distinct editing complexes exist. Complex I (19S) consists of gRNA, TUTase, RNA ligase and chimera-forming activity. Complex II (35-40S) is composed of gRNA, preedited mRNA, RNA ligase and chimera-forming activity. These studies provide the first evidence that editing occurs in a multicomponent complex. The possible roles of complex I, complex II and RNA ligase in editing are discussed.
本研究的目的是鉴定参与动基体线粒体mRNA编辑的多组分复合物。将布氏锥虫的线粒体提取物在10%-30%的甘油梯度上进行分级分离,并检测可能参与编辑的RNA和活性,包括编辑前的mRNA、引导RNA(gRNA)、内切核酸酶、末端尿苷酰转移酶(TUTase)、RNA连接酶和gRNA-mRNA嵌合体形成活性。这些实验表明存在两种不同的编辑复合物。复合物I(19S)由gRNA、TUTase、RNA连接酶和嵌合体形成活性组成。复合物II(35-40S)由gRNA、编辑前的mRNA、RNA连接酶和嵌合体形成活性组成。这些研究提供了第一个证据,证明编辑发生在多组分复合物中。讨论了复合物I、复合物II和RNA连接酶在编辑中的可能作用。