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火鸡转录因子Pit-1/GHF-1在体外可激活火鸡催乳素和生长激素基因启动子,但在体内的催乳细胞中无法检测到。

The turkey transcription factor Pit-1/GHF-1 can activate the turkey prolactin and growth hormone gene promoters in vitro but is not detectable in lactotrophs in vivo.

作者信息

Weatherly K L, Ramesh R, Strange H, Waite K L, Storrie B, Proudman J A, Wong E A

机构信息

Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, Virginia 24061-0306, USA.

出版信息

Gen Comp Endocrinol. 2001 Sep;123(3):244-53. doi: 10.1006/gcen.2001.7680.

Abstract

The transcription factor Pit-1/GHF-1 plays an important role in regulating the prolactin (Prl) and growth hormone (GH) genes in mammals. In this study, the role that Pit-1 plays in regulating the prolactin and growth hormone genes in avian species was examined by cotransfection assays and immunofluorescence staining of pituitary sections. In cotransfection assays, turkey Pit-1 activated the turkey Prl, turkey GH, and rat Prl promoters 3.8-, 3.7-, and 12.5-fold, respectively. This activation was comparable to rat Pit-1 activation of these same promoters. A point mutation in the turkey Pit-1 cDNA, which changed leu-219 to ser-219, resulted in a 2-, 2-, and 10-fold reduction in the activation of the turkey Prl, turkey GH, and rat Prl promoters, respectively. Unexpectedly, coexpression of tPit-1 (leu-219) and tPit-1(ser-219) activated turkey Prl and rat Prl promoters 9.4- and 35.9-fold, respectively, but had no effect on the turkey GH promoter. Dual-label immunofluorescence analysis of turkey pituitary sections revealed that Pit-1 was not detectable in prolactin-staining cells but was detectable in GH-staining cells. Taken together, these data indicate that in the domestic turkey, Pit-1 can activate the turkey Prl promoter in vitro, but does not appear to play a role in regulating Prl gene expression in vivo. Pit-1, however, still likely plays a role in regulating GH gene expression.

摘要

转录因子Pit-1/GHF-1在调节哺乳动物的催乳素(Prl)和生长激素(GH)基因方面发挥着重要作用。在本研究中,通过共转染实验和垂体切片的免疫荧光染色,研究了Pit-1在调节禽类催乳素和生长激素基因中的作用。在共转染实验中,火鸡Pit-1分别使火鸡Prl、火鸡GH和大鼠Prl启动子激活了3.8倍、3.7倍和12.5倍。这种激活作用与大鼠Pit-1对相同启动子的激活作用相当。火鸡Pit-1 cDNA中的一个点突变将亮氨酸-219变为丝氨酸-219,导致火鸡Prl、火鸡GH和大鼠Prl启动子的激活分别降低了2倍、2倍和10倍。出乎意料的是,tPit-1(亮氨酸-219)和tPit-1(丝氨酸-219)的共表达分别使火鸡Prl和大鼠Prl启动子激活了9.4倍和35.9倍,但对火鸡GH启动子没有影响。对火鸡垂体切片的双标记免疫荧光分析显示,在催乳素染色细胞中未检测到Pit-1,但在GH染色细胞中可检测到。综上所述,这些数据表明,在家养火鸡中,Pit-1在体外可激活火鸡Prl启动子,但在体内调节Prl基因表达方面似乎不起作用。然而,Pit-1可能仍在调节GH基因表达中发挥作用。

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