Sirito M, Lin Q, Maity T, Sawadogo M
Department of Molecular Genetics, University of Texas, M.D. Anderson Cancer Center, Houston 77030.
Nucleic Acids Res. 1994 Feb 11;22(3):427-33. doi: 10.1093/nar/22.3.427.
USF is a helix-loop-helix transcription factor that, like Myc, recognizes the DNA binding motif CACGTG. Two different forms of USF, characterized by apparent molecular weights of 43,000 and 44,000, were originally identified in HeLa cells by biochemical analysis. Clones for the 43-kDa USF were first characterized, but only partial clones for the human 44-kDa USF (USF2, or FIP) have been reported. Here we describe a complete cDNA for the 44-kDa USF from murine cells. Analysis of this clone has revealed that the various USF family members are quite divergent in their N-terminal amino acid sequences, while a high degree of conservation characterizes their dimerization and DNA-binding domains. Interestingly, the 3' noncoding region of the 44-kDa USF cDNAs displayed an unusual degree of conservation between human and mouse. In vitro transcription/translation experiments indicated a possible role for this region in translation regulation. Alternative splicing forms of the 44-kDa USF messages exist in both mouse and human. Examination of the tissue and cell-type distribution of USF by Northern blot and gel retardation assays revealed that while expression of both the 43- and 44-kDa USF species is ubiquitous, different ratios of USF homo- and heterodimers are found in different cells.
USF是一种螺旋-环-螺旋转录因子,与Myc一样,能识别DNA结合基序CACGTG。通过生化分析,最初在HeLa细胞中鉴定出两种不同形式的USF,其表观分子量分别为43000和44000。首先对43 kDa的USF克隆进行了表征,但关于人类44 kDa的USF(USF2或FIP)仅报道了部分克隆。在此,我们描述了来自鼠细胞的44 kDa USF的完整cDNA。对该克隆的分析表明,USF家族的各个成员在其N端氨基酸序列上差异很大,而其二聚化和DNA结合域则具有高度保守性。有趣的是,44 kDa USF cDNA的3'非编码区在人和小鼠之间表现出异常高的保守性。体外转录/翻译实验表明该区域在翻译调控中可能发挥作用。在小鼠和人类中都存在44 kDa USF信使的可变剪接形式。通过Northern印迹和凝胶阻滞试验检测USF的组织和细胞类型分布,结果显示,虽然43 kDa和44 kDa的USF均普遍表达,但在不同细胞中发现了不同比例的USF同二聚体和异二聚体。