van den Boom Vincent, Citterio Elisabetta, Hoogstraten Deborah, Zotter Angelika, Egly Jean-Marc, van Cappellen Wiggert A, Hoeijmakers Jan H J, Houtsmuller Adriaan B, Vermeulen Wim
Department of Cell Biology and Genetics, Erasmus MC, P.O. Box 1738, 3000 DR Rotterdam, Netherlands.
J Cell Biol. 2004 Jul 5;166(1):27-36. doi: 10.1083/jcb.200401056. Epub 2004 Jun 28.
The Cockayne syndrome B (CSB) protein is essential for transcription-coupled DNA repair (TCR), which is dependent on RNA polymerase II elongation. TCR is required to quickly remove the cytotoxic transcription-blocking DNA lesions. Functional GFP-tagged CSB, expressed at physiological levels, was homogeneously dispersed throughout the nucleoplasm in addition to bright nuclear foci and nucleolar accumulation. Photobleaching studies showed that GFP-CSB, as part of a high molecular weight complex, transiently interacts with the transcription machinery. Upon (DNA damage-induced) transcription arrest CSB binding these interactions are prolonged, most likely reflecting actual engagement of CSB in TCR. These findings are consistent with a model in which CSB monitors progression of transcription by regularly probing elongation complexes and becomes more tightly associated to these complexes when TCR is active.
科凯恩综合征B(CSB)蛋白对于转录偶联DNA修复(TCR)至关重要,而TCR依赖于RNA聚合酶II的延伸。TCR是快速去除细胞毒性转录阻断性DNA损伤所必需的。在生理水平表达的功能性绿色荧光蛋白(GFP)标记的CSB,除了明亮的核灶和核仁积累外,还均匀地分散在整个核质中。光漂白研究表明,作为高分子量复合物的一部分,GFP-CSB与转录机制瞬时相互作用。在(DNA损伤诱导的)转录停滞时,CSB的结合这些相互作用会延长,这很可能反映了CSB在TCR中的实际参与。这些发现与一个模型一致,即CSB通过定期探测延伸复合物来监测转录进程,并在TCR激活时与这些复合物更紧密地结合。