Safrany G, Tanaka N, Kishimoto T, Ishikawa Y, Kato H, Kominami R, Muramatsu M
Department of Biochemistry, University of Tokyo Faculty of Medicine, Japan.
Mol Cell Biol. 1989 Jan;9(1):349-53. doi: 10.1128/mcb.9.1.349-353.1989.
Mammalian ribosomal DNA (rDNA) transcription has a certain species specificity such that, both in vivo and in vitro, human rDNA cannot be transcribed by mouse machinery and vice versa. This is due to a species-dependent transcription factor, TFID (Y. Mishima, I. Financsek, R. Kominami, and M. Muramatsu, Nucleic Acids Res. 10:6659-6670, 1982). On the basis of the information obtained from 5' and 3' substitution mutants, we prepared a chimeric gene in which the mouse sequence from positions -32 to -14 was inserted into the corresponding location of the human rDNA promoter. The chimeric gene could be transcribed by mouse extracts nearly as efficiently as the wild-type mouse promoter. The chimeric gene could also sequester transcription factor TFID at an efficiency similar to that for the mouse promoter. Partially purified mouse TFID that could not protect the human rDNA promoter against DNase I produced a clear footprint on this chimeric gene that was similar to that on mouse rDNA promoter. The basic structure of the mouse rDNA core promoter is discussed in relation to the interaction with TFID.
哺乳动物核糖体DNA(rDNA)转录具有一定的物种特异性,以至于在体内和体外,人类rDNA都不能被小鼠的转录机制转录,反之亦然。这是由于一种物种依赖性转录因子TFID(Y. 三岛、I. 菲南塞克、R. 小南和M. 村松,《核酸研究》10:6659 - 6670,1982年)。基于从5'和3'取代突变体获得的信息,我们制备了一个嵌合基因,其中将小鼠 - 32至 - 14位的序列插入到人类rDNA启动子的相应位置。该嵌合基因能够被小鼠提取物转录,效率几乎与野生型小鼠启动子相同。该嵌合基因还能够以与小鼠启动子相似的效率隔离转录因子TFID。不能保护人类rDNA启动子免受DNase I作用的部分纯化的小鼠TFID,在该嵌合基因上产生了一个清晰的足迹,与在小鼠rDNA启动子上的足迹相似。本文讨论了小鼠rDNA核心启动子的基本结构与TFID相互作用的关系。