Kropotov A V, Tomilin N V
Institute of cytology of the Russian Academy of Sciences, St. Petersburg, Russian Federation.
FEBS Lett. 1996 May 13;386(1):43-6. doi: 10.1016/0014-5793(96)00353-5.
Internal promoters of some genes transcribed by RNA polymerase III (e.g. tRNA genes, adenovirus VA1 RNA gene, human retroposons of the Alu family) contain a conserved sequence element, B-box, interacting with basal transcription factor TFIIIC2 which initiates assembly of the full transcription complex on the genes, and which represents the major determinant of the efficiency of their expression. In this study we have identified in human nuclear extracts a protein which interacts with VA1 B-box DNA and forms a high-affinity complex which is very stable after the addition of a large excess of competitor DNA. Unlike TFIIIC2, the B-box-binding activity of the B-box-binding protein is found to be decreased in adenovirus 5-transformed human cells. In these cells (line 293) increased transcription of VA1 and tRNA genes in vivo and in vitro was previously detected by other workers. Our results suggest that besides TFIIIC2, an additional B-box-binding protein factor may be involved in the regulation of expression of the RNA polymerase III-transcribed genes.
一些由RNA聚合酶III转录的基因(如tRNA基因、腺病毒VA1 RNA基因、Alu家族的人类反转座子)的内部启动子含有一个保守的序列元件,即B框,它与基础转录因子TFIIIC2相互作用,TFIIIC2启动完整转录复合体在这些基因上的组装,并且是其表达效率的主要决定因素。在本研究中,我们在人核提取物中鉴定出一种与VA1 B框DNA相互作用并形成高亲和力复合体的蛋白质,在加入大量过量的竞争DNA后,该复合体非常稳定。与TFIIIC2不同,B框结合蛋白的B框结合活性在腺病毒5转化的人细胞中降低。在这些细胞(293细胞系)中,其他研究人员之前已检测到体内和体外VA1和tRNA基因转录增加。我们的结果表明,除了TFIIIC2外,另一种B框结合蛋白因子可能参与RNA聚合酶III转录基因表达的调控。