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过氧化物酶体增殖物激活受体(PPAR)γ的多种变体在大西洋鲑(Salmo salar)的肝脏中表达。

Multiple variants of the peroxisome proliferator-activated receptor (PPAR) gamma are expressed in the liver of atlantic salmon (Salmo salar).

作者信息

Andersen O, Eijsink V G, Thomassen M

机构信息

Akvaforsk, Institute of Aquaculture Research Ltd., PO Box 5010, N-1432, Aas, Norway.

出版信息

Gene. 2000 Sep 19;255(2):411-8. doi: 10.1016/s0378-1119(00)00350-4.

Abstract

A full-length cDNA encoding the peroxisome proliferator-activated receptor (PPAR) has for the first time been characterized from a fish species. The Atlantic salmon PPARgamma cDNA of 2528 nucleotides (nt) was amplified from liver mRNA by reverse transcription (RT)-polymerase chain reaction (PCR). The deduced protein of 544 amino acids (aa) shares approximately 47% overall sequence identity with mammalian PPARgamma. The N-terminal A/B region contains a repeated decapeptide motif and shows a low homology with other PPARs. In contrast, the central DNA-binding domain (DBD) and the C-terminal ligand-binding domain (LBD) show a high sequence identity to mammalian and Xenopus PPARgamma. The salmon PPARgamma LBD contains nine additional residues in a flexible loop that might affect ligand binding. Northern blot analysis of salmon liver RNA revealed a prominent transcript of about 1.7 kilo bases (kb), in addition to several mRNA species of about 2.4-2.6kb, which is consistent with the presence of multiple putative polyadenylation sites in the 3' untranslated region (UTR) of the 2528nt long PPARgamma cDNA. Two additional PPARgamma cDNAs of 1719 and 2357nt were then isolated. The 2357nt long transcript encodes full-length PPARgamma and seems to be ubiquitously expressed in salmon, whereas the liver-specific transcript of 1719nt encodes a truncated variant of PPARgamma. The truncated form lacks 39 C-terminal residues including the conserved activation function-2 (AF-2) motif, known to be associated with crucial cofactors. Three-dimensional modelling studies indicated that the C-terminal truncation would result in important alterations of the ligand-binding pocket. The presence of a truncated form with drastic changes in both ligand- and cofactor-binding sites is likely to modulate PPARgamma activity in salmon liver.

摘要

首次从一种鱼类中鉴定出编码过氧化物酶体增殖物激活受体(PPAR)的全长cDNA。通过逆转录(RT)-聚合酶链反应(PCR)从肝脏mRNA中扩增出2528个核苷酸(nt)的大西洋鲑PPARγ cDNA。推导的含544个氨基酸(aa)的蛋白质与哺乳动物PPARγ的总体序列同一性约为47%。N端A/B区域包含一个重复的十肽基序,与其他PPARs的同源性较低。相比之下,中央DNA结合结构域(DBD)和C端配体结合结构域(LBD)与哺乳动物和非洲爪蟾PPARγ具有高度的序列同一性。鲑鱼PPARγ LBD在一个柔性环中含有另外九个残基,这可能会影响配体结合。对鲑鱼肝脏RNA的Northern印迹分析显示,除了几种约2.4 - 2.6kb的mRNA种类外,还有一个约1.7千碱基(kb)的突出转录本,这与2528nt长的PPARγ cDNA的3'非翻译区(UTR)中存在多个假定的聚腺苷酸化位点一致。随后分离出另外两个分别为1719和2357nt的PPARγ cDNA。2357nt长的转录本编码全长PPARγ,似乎在鲑鱼中普遍表达,而1719nt的肝脏特异性转录本编码PPARγ的截短变体。截短形式缺少39个C端残基,包括已知与关键辅因子相关的保守激活功能-2(AF-2)基序。三维建模研究表明,C端截短将导致配体结合口袋的重要改变。在配体和辅因子结合位点都有剧烈变化的截短形式的存在可能会调节鲑鱼肝脏中PPARγ的活性。

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