Landrieux Emilie, Alic Nazif, Ducrot Cécile, Acker Joël, Riva Michel, Carles Christophe
CEA/Saclay, Laboratoire de Transcription des Gènes, Service de Biochimie et de Génétique Moléculaire, Gif sur Yvette, France.
EMBO J. 2006 Jan 11;25(1):118-28. doi: 10.1038/sj.emboj.7600915. Epub 2005 Dec 15.
While initiation of transcription by RNA polymerase III (Pol III) has been thoroughly investigated, molecular mechanisms driving transcription termination remain poorly understood. Here we describe how the characterization of the in vitro transcriptional properties of a Pol III variant (Pol IIIdelta), lacking the C11, C37, and C53 subunits, revealed crucial information about the mechanisms of Pol III termination and reinitiation. The specific requirement for the C37-C53 complex in terminator recognition was determined. This complex was demonstrated to slow down elongation by the enzyme, adding to the evidence implicating the elongation rate as a critical determinant of correct terminator recognition. In addition, the presence of the C37-C53 complex required the simultaneous addition of C11 to Pol IIIdelta for the enzyme to reinitiate after the first round of transcription, thus uncovering a role for polymerase subunits in the facilitated recycling process. Interestingly, we demonstrated that the role of C11 in recycling was independent of its role in RNA cleavage. The data presented allowed us to propose a model of Pol III termination and its links to reinitiation.
虽然RNA聚合酶III(Pol III)引发转录的过程已得到充分研究,但驱动转录终止的分子机制仍知之甚少。在此,我们描述了对缺乏C11、C37和C53亚基的Pol III变体(Pol IIIdelta)体外转录特性的表征,如何揭示了有关Pol III终止和重新起始机制的关键信息。确定了C37 - C53复合物在终止子识别中的特定要求。已证明该复合物会减缓酶的延伸速度,这进一步证明延伸速率是正确识别终止子的关键决定因素。此外,C37 - C53复合物的存在要求在第一轮转录后,同时向Pol IIIdelta添加C11,以使酶能够重新起始,从而揭示了聚合酶亚基在促进循环过程中的作用。有趣的是,我们证明了C11在循环中的作用与其在RNA切割中的作用无关。所呈现的数据使我们能够提出一个Pol III终止模型及其与重新起始的联系。