Scotto K W, Kaulen H, Roeder R G
Rockefeller University, New York, New York 10021.
Genes Dev. 1989 May;3(5):651-62. doi: 10.1101/gad.3.5.651.
Expression of the positively acting 5S gene-specific transcription factor, TFIIIA, is regulated during development, with highest levels of mRNA and protein occurring during oogenesis. By analysis of TFIIIA promoter mutants microinjected into late stage Xenopus oocytes, we have determined DNA sequences required for the transcription of this gene and we have identified proteins that bind to these regulatory sequences. A negative element lies between positions -306 and -289. Three positive-acting sequences are located between positions -289 and -253, -250 and -173, and -144 and -101. Gel shift analyses of TFIIIA promoter fragments incubated with Xenopus oocyte extracts have identified two DNA-protein complexes. One complex, designated B1, requires sequences within the promoter region extending from -271 to -253 while the second complex, designated B2, involves promoter sequences from -235 to -221. The protein involved in formation of the B1 complex has been found to be related to the human adenovirus major late transcription factor, USF.
正向作用的5S基因特异性转录因子TFIIIA的表达在发育过程中受到调控，在卵子发生过程中mRNA和蛋白质水平最高。通过对显微注射到非洲爪蟾晚期卵母细胞中的TFIIIA启动子突变体进行分析，我们确定了该基因转录所需的DNA序列，并鉴定了与这些调控序列结合的蛋白质。一个负调控元件位于-306至-289位之间。三个正向作用序列分别位于-289至-253位、-250至-173位以及-144至-101位之间。用非洲爪蟾卵母细胞提取物孵育TFIIIA启动子片段进行凝胶迁移分析，鉴定出两种DNA-蛋白质复合物。一种复合物命名为B1，需要启动子区域内从-271至-253位的序列，而第二种复合物命名为B2，涉及从-235至-221位的启动子序列。已发现参与形成B1复合物的蛋白质与人类腺病毒主要晚期转录因子USF相关。
