Mocikat R, Falkner F G, Mertz R, Zachau H G
Nucleic Acids Res. 1986 Nov 25;14(22):8829-44. doi: 10.1093/nar/14.22.8829.
The decanucleotide sequence (dc) TNATTTGCAT is an upstream regulatory sequence of immunoglobulin genes and occurs also upstream of certain other eukaryotic and prokaryotic genes (compiled in the accompanying paper). We now investigated the binding of proteins from nuclear extracts of a number of cell types and organisms to the dc sequence using a sensitive gel electrophoretic DNA binding assay. Binding studies with specifically designed oligonucleotides led to the following conclusions: the central T of the dc sequence can be altered with only a slight decrease of protein binding activity: the sequences in the neighborhood of dc have a positive or negative effect on the efficiency of protein binding; C-rich sequences which occur in many K chain promoters have a protein binding activity independent of dc; the dc binding protein(s) of human lymphoid cells elute from a Sephadex column in the 30.000-60.000 molecular weight range; dc binding proteins were found in nuclear extracts of lymphoid as well as non-lymphoid human and murine cell lines, of Xenopus oocytes, and of yeast cells. The finding of dc binding proteins in a wide variety of different organisms and the occurrence of dc-related sequences in the regulatory regions of several gene families point to a general role in the transcriptional regulation of the respective genes.
十聚体序列(dc)TNATTTGCAT是免疫球蛋白基因的上游调控序列,在某些其他真核和原核基因的上游也有出现(在随附论文中汇编)。我们现在使用灵敏的凝胶电泳DNA结合分析法,研究了多种细胞类型和生物体的核提取物中的蛋白质与dc序列的结合情况。对经过特殊设计的寡核苷酸进行的结合研究得出了以下结论:dc序列的中心T可以改变,而蛋白质结合活性仅略有下降;dc附近的序列对蛋白质结合效率有正向或负向影响;许多K链启动子中出现的富含C的序列具有独立于dc的蛋白质结合活性;人淋巴细胞的dc结合蛋白在分子量30,000 - 60,000范围内从Sephadex柱上洗脱下来;在人及小鼠的淋巴细胞和非淋巴细胞系、非洲爪蟾卵母细胞以及酵母细胞的核提取物中都发现了dc结合蛋白。在多种不同生物体中发现dc结合蛋白以及在几个基因家族的调控区域中出现与dc相关的序列,这表明其在各自基因的转录调控中具有普遍作用。