Kokubo T, Gong D W, Roeder R G, Horikoshi M, Nakatani Y
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5896-900. doi: 10.1073/pnas.90.13.5896.
Transcription initiation factor TFIID is a multimeric protein complex that plays a central role in transcriptional regulation by facilitating promoter responses to various activators. cDNAs encoding the 110-kDa subunit of Drosophila TFIID (p110) were isolated with a degenerate oligodeoxynucleotide probe based on an amino acid sequence of the purified protein. The entire cDNA sequence contains an open reading frame encoding a 921-amino acid polypeptide with a calculated molecular mass of 99,337 Da. The recombinant protein expressed in Sf9 cells via a baculovirus vector interacts directly with the 230-kDa subunit of TFIID (p230). Together with the previous observation that the TATA box-binding subunit of TFIID (TFIID tau or TBP) interacts directly with only p230 among the TFIID subunits, this result suggests that p110 forms a complex with TFIID tau via p230. A binding study using various p230 mutants indicated that both p110 and TFIID tau interact with the N-terminal 352-amino acid portion of p230, suggesting a functional communication between p110 and TFIID tau via p230 interactions.
转录起始因子TFIID是一种多聚体蛋白复合物,通过促进启动子对各种激活因子的反应在转录调控中发挥核心作用。基于纯化蛋白的氨基酸序列,用简并寡脱氧核苷酸探针分离出编码果蝇TFIID 110 kDa亚基(p110)的cDNA。整个cDNA序列包含一个开放阅读框,编码一个921个氨基酸的多肽,计算分子量为99,337 Da。通过杆状病毒载体在Sf9细胞中表达的重组蛋白直接与TFIID的230 kDa亚基(p230)相互作用。连同之前观察到的TFIID的TATA盒结合亚基(TFIID tau或TBP)在TFIID亚基中仅直接与p230相互作用这一结果,该结果表明p110通过p230与TFIID tau形成复合物。使用各种p230突变体的结合研究表明,p110和TFIID tau都与p230的N端352个氨基酸部分相互作用,这表明p110和TFIID tau通过p230相互作用存在功能联系。