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鲑鱼肝细胞核因子1:cDNA序列、进化、组织特异性及与鲑鱼血清白蛋白启动子的结合

Salmon HNF1: cDNA sequence, evolution, tissue specificity and binding to the salmon serum albumin promoter.

作者信息

Deryckere F, Byrnes L, Wagner A, McMorrow T, Gannon F

机构信息

National Diagnostics Centre/BioResearch Ireland, Department of Microbiology, University College Galway.

出版信息

J Mol Biol. 1995 Mar 17;247(1):1-10. doi: 10.1006/jmbi.1994.0115.

Abstract

cDNA clones coding for the transcription factor HNF1 have been isolated from Atlantic salmon (Salmo salar L.). The 559 amino acid residue long encoded protein shows high conservation, with respect to other species, of the domains necessary for DNA-binding: the HNF1 atypical homeodomain, the POU related sequence and the dimerisation domain. Alignment with rat HNF1 protein reveals that the transcription activation domains ADI and ADIII are relatively conserved in the fish sequence whereas ADII is not. Phylogenetic analysis indicates that higher vertebrate HNF1s and the related variant HNF1s (vHNF1s) are more closely related to each other than any of them is to Salmon HNF1, suggesting that the duplication event from which HNF1 and vHNF1 genes arose occurred after the divergence of the tetrapod and teleost ancestors. Northern blot analysis show a single transcript, of about 2.6 kb, which is not exclusive to liver but is also present in intestine, kidney and spleen. Using polymerase chain reaction (PCR) we have isolated the salmon albumin gene promoter which contains, upstream of the TATA box, a potential binding site for HNF1. The salmon HNF1 protein synthesized by in vitro transcription-translation of the full-length cDNA is able to bind specifically with equivalent affinities to either the rat or salmon albumin promoter.

摘要

编码转录因子HNF1的cDNA克隆已从大西洋鲑(Salmo salar L.)中分离出来。编码的蛋白质有559个氨基酸残基,与其他物种相比,其DNA结合所需结构域具有高度保守性:HNF1非典型同源结构域、POU相关序列和二聚化结构域。与大鼠HNF1蛋白比对发现,转录激活结构域ADI和ADIII在鱼类序列中相对保守,而ADII则不然。系统发育分析表明,高等脊椎动物的HNF1和相关变体HNF1(vHNF1)彼此之间的关系比它们与鲑鱼HNF1的关系更为密切;这表明HNF1和vHNF1基因产生的复制事件发生在四足动物和硬骨鱼祖先分化之后。Northern印迹分析显示有一个约2.6kb的单一转录本,它并非肝脏所特有,在肠道、肾脏和脾脏中也存在。利用聚合酶链反应(PCR),我们分离出了鲑鱼白蛋白基因启动子,该启动子在TATA框上游含有一个HNF1的潜在结合位点。通过对全长cDNA进行体外转录-翻译合成的鲑鱼HNF1蛋白能够以相同亲和力与大鼠或鲑鱼白蛋白启动子特异性结合。

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