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在转基因小鼠中,精子细胞中睾丸激素敏感性脂肪酶的表达受一个短启动子调控。

Testis hormone-sensitive lipase expression in spermatids is governed by a short promoter in transgenic mice.

作者信息

Blaise R, Guillaudeux T, Tavernier G, Daegelen D, Evrard B, Mairal A, Holm C, Jégou B, Langin D

机构信息

INSERM Unit 317, Institut Louis Bugnard, Université Paul Sabatier, Hôpital Rangueil, F-31403 Toulouse Cedex 4, France.

出版信息

J Biol Chem. 2001 Feb 16;276(7):5109-15. doi: 10.1074/jbc.M009103200. Epub 2000 Nov 13.

Abstract

A testicular form of hormone-sensitive lipase (HSL(tes)), a triacylglycerol lipase, and cholesterol esterase, is expressed in male germ cells. Northern blot analysis showed HSL(tes) mRNA expression in early spermatids. Immunolocalization of the protein in human and rodent seminiferous tubules indicated that the highest level of expression occurred in elongated spermatids. We have previously shown that 0.5 kilobase pairs of the human HSL(tes) promoter directs testis-specific expression of a chloramphenicol acetyltransferase reporter gene in transgenic mice and determined regions binding nuclear proteins expressed in testis but not in liver (Blaise, R., Grober, J., Rouet, P., Tavernier, G., Daegelen, D., and Langin, D. (1999) J. Biol. Chem. 274, 9327-9334). Mutation of a SRY/Sox-binding site in one of the regions did not impair in vivo testis-specific expression of the reporter gene. Further transgenic analyses established that 95 base pairs upstream of the transcription start site were sufficient for correct testis expression. In gel retardation assays using early spermatid nuclear extracts, a germ cell-specific DNA-protein interaction was mapped between -46 and -29 base pairs. The DNA binding nuclear protein showed properties of zinc finger transcription factors. Mutation of the region abolished reporter gene activity in transgenic mice, showing that it is necessary for testis expression of HSL(tes).

摘要

激素敏感脂肪酶的睾丸形式(HSL(tes)),一种三酰甘油脂肪酶和胆固醇酯酶,在雄性生殖细胞中表达。Northern印迹分析显示HSL(tes) mRNA在早期精子细胞中表达。该蛋白在人和啮齿动物曲细精管中的免疫定位表明,最高表达水平出现在延长型精子细胞中。我们之前已经表明,人HSL(tes)启动子的0.5千碱基对可指导氯霉素乙酰转移酶报告基因在转基因小鼠中的睾丸特异性表达,并确定了与睾丸而非肝脏中表达的核蛋白结合的区域(布莱斯,R.,格罗贝尔,J.,鲁埃,P.,塔维尼耶,G.,代盖伦,D.,和兰金,D.(1999年)《生物化学杂志》274,9327 - 9334)。其中一个区域中SRY/Sox结合位点发生突变并不损害报告基因在体内的睾丸特异性表达。进一步的转基因分析确定转录起始位点上游95个碱基对足以实现正确的睾丸表达。在使用早期精子细胞核提取物的凝胶阻滞分析中,一种生殖细胞特异性的DNA - 蛋白质相互作用被定位在 - 46至 - 29个碱基对之间。该DNA结合核蛋白显示出锌指转录因子的特性。该区域发生突变消除了转基因小鼠中报告基因的活性,表明它对于HSL(tes)的睾丸表达是必需的。

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