Weinrich S L, Meister A, Rutter W J
Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0534.
Mol Cell Biol. 1991 Oct;11(10):4985-97. doi: 10.1128/mcb.11.10.4985-4997.1991.
Exocrine pancreas (XP) enhancers, which contain a conserved core sequence, are active only in XP cells. A core enhancer-binding activity also appears to be restricted to XP nuclei. Here we describe the properties of a factor, purified approximately 100,000-fold from pancreas nuclei, which displays core enhancer-binding activity. It is not identical to previously characterized factors and is termed exocrine pancreas transcription factor 1 (XPF-1). In the highly purified preparation, only a single major protein of 60 kDa was detected by silver staining on sodium dodecyl sulfate-gels and by UV cross-linking. XPF-1 binds to the core enhancer of all tested XP genes and not to a mutant sequence which is inactive in vivo. High-affinity binding sites are bipartite. The results of competition binding and UV-cross-linking assays suggest that XPF-1 interacts with both motifs. XPF-1 selectively stimulates transcription of core enhancer templates in an in vitro transcription system. We hypothesize that XPF-1 plays a role in activation of the transcription of XP-specific genes.
外分泌胰腺(XP)增强子含有保守的核心序列,仅在XP细胞中具有活性。一种核心增强子结合活性似乎也仅限于XP细胞核。在此,我们描述了一种从胰腺细胞核中纯化了约10万倍的因子的特性,该因子具有核心增强子结合活性。它与先前鉴定的因子不同,被称为外分泌胰腺转录因子1(XPF-1)。在高度纯化的制剂中,通过十二烷基硫酸钠凝胶上的银染和紫外线交联,在十二烷基硫酸钠凝胶上仅检测到一种60 kDa的主要蛋白质。XPF-1与所有测试的XP基因的核心增强子结合,而不与体内无活性的突变序列结合。高亲和力结合位点是二分的。竞争结合和紫外线交联试验的结果表明,XPF-1与两个基序相互作用。XPF-1在体外转录系统中选择性地刺激核心增强子模板的转录。我们假设XPF-1在XP特异性基因转录的激活中起作用。