Huang W T, Gong H Y, Lin C J, Weng C F, Chen M H, Wu J L
Institute of Zoology, Academia Sinica, Nankang, Taipei, Taiwan, 11529, Republic of China.
Biochem Biophys Res Commun. 2001 Nov 9;288(4):833-40. doi: 10.1006/bbrc.2001.5856.
Hepatocyte nuclear factors (HNFs) are upstream regulators of many liver-specific genes and are involved in many cellular functions in the body, but their existence, expression, and function in gonads are still poorly understood. Here we report on the first cloning of partial cDNAs of HNF-1alpha and -1beta and full HNF-3beta cDNA from a tilapia (Oreochromis mossambicus) liver cDNA library. The deduced amino acid sequence of tilapia HNF-3beta has a 90 to 96% identity with those of other fishes (dwarf gourami, medaka, and zebrafish), 74% with mammals (human, rat, and mouse), and 82% with Xenopus. RT-PCR detected IGF-I and -II and HNF-1alpha, -1beta, and -3beta in both liver and gonads and the identity of the PCR fragments was confirmed by PCR hybridization. Immunoprecipitation and Western blotting also detected all three HNF proteins in both liver and gonads. Expression of HNFs in the gonads of the tilapia suggests that multi-HNFs may form a cascade to regulate gonadal physiology in the bony fish.
肝细胞核因子(HNFs)是许多肝脏特异性基因的上游调节因子,参与身体的多种细胞功能,但它们在性腺中的存在、表达和功能仍知之甚少。在此,我们报道了首次从罗非鱼(莫桑比克罗非鱼)肝脏cDNA文库中克隆出HNF-1α和-1β的部分cDNA以及完整的HNF-3β cDNA。罗非鱼HNF-3β推导的氨基酸序列与其他鱼类(侏儒 Gourami、青鳉和斑马鱼)的序列具有90%至96%的同一性,与哺乳动物(人、大鼠和小鼠)的序列同一性为74%,与非洲爪蟾的序列同一性为82%。RT-PCR在肝脏和性腺中均检测到IGF-I和-II以及HNF-1α、-1β和-3β,PCR片段的同一性通过PCR杂交得以证实。免疫沉淀和蛋白质印迹也在肝脏和性腺中检测到了所有三种HNF蛋白。罗非鱼性腺中HNFs的表达表明,多种HNFs可能形成一个级联反应来调节硬骨鱼的性腺生理。