van Wijnen A J, Massung R F, Stein J L, Stein G S
Department of Cell Biology, University of Massachusetts Medical School, Worcester 01655.
Biochemistry. 1988 Aug 23;27(17):6534-41. doi: 10.1021/bi00417a051.
Vertebrate histone gene promoters in many cases contain an upstream element, 5'dCCAAT, that has been implicated in modulating the efficiency of transcription of a broad spectrum of genes. We have previously isolated a nuclear factor (HiNF-B) that binds specifically to the CCAAT element of a cell cycle regulated human H1 histone gene. This factor shows similarities with other CCAAT box binding proteins in that it recognizes the same sequence but shows a distinct chromatographic behavior. In the present study, we have employed the gel retardation assay to demonstrate that HiNF-B is a cell cycle independent DNA binding protein that is conserved in both human and mouse cells. Using a series of reconstitution experiments with partially purified HiNF-B fractions, we show that this factor requires association of at least two components for site-specific binding. The composite structure of HiNF-B suggests that binding of at least some CCAAT elements in vertebrates may require cooperative interaction of CCAAT box binding proteins with other factors.
在许多情况下,脊椎动物组蛋白基因启动子包含一个上游元件,即5'dCCAAT,它与调节多种基因的转录效率有关。我们之前分离出一种核因子(HiNF-B),它能特异性结合细胞周期调控的人类H1组蛋白基因的CCAAT元件。该因子与其他CCAAT盒结合蛋白相似,因为它识别相同的序列,但表现出独特的色谱行为。在本研究中,我们采用凝胶阻滞试验证明HiNF-B是一种细胞周期非依赖性DNA结合蛋白,在人类和小鼠细胞中均保守。通过一系列对部分纯化的HiNF-B组分的重组实验,我们表明该因子需要至少两种组分结合才能实现位点特异性结合。HiNF-B的复合结构表明,脊椎动物中至少某些CCAAT元件的结合可能需要CCAAT盒结合蛋白与其他因子的协同相互作用。