The distal regulatory region of the human involucrin promoter is required for expression in epidermis
The distal regulatory region of the human involucrin promoter is required for expression in epidermis
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DOI：
10.1074/jbc.273.46.30460
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发表时间：
1998-11-13
影响因子：
4.8
通讯作者：
Eckert, RL
中科院分区：
文献类型：
--
作者：
Crish, JF;Zaim, TM;Eckert, RL
人总蛋白(HINV)是角质形成细胞角化包膜的前体，它特异地表达于复层鳞状上皮的超基底层。启动子远端(DRR)和近端调控区(PRR)是体外最佳表达所必需的(Welter，J，F，，Crish，J.F.，Agarwal，C.，和Eckert，R.L，(1995)J.Biol)。化学。12614-12622；以及Banks，E.B.，Crish，J.F.，Welter，J.F.，和Eckert，R.L.(1998年)生物化学。J.331、61-68)。我们现在公布这些区域的完整序列，并评估它们驱动体内转录的能力。含有5000或2473碱基对上游调控区的转基因在复层表面上皮细胞中驱动组织和分化的适当表达，相反，含有1953、1333、986或41碱基对上游调控区的转基因在表面上皮细胞中不表达，这表明DRR(核苷酸-2474/-1953)的缺失导致了表达的丧失。将分离的DRR区域直接与hINV最小启动子融合，可恢复表面上皮细胞的表达。转录基因下游的序列不需要适当的表达。-1953/-41片段影响依赖分化的表达模式。-986/-41区域包括PRR，驱动内上皮细胞的表达。
Human involucrin (hINV) is a precursor of the keratinocyte cornified envelope that is specifically expressed in the suprabasal layers of stratifying squamous epithelia. The promoter distal (DRR) and proximal regulatory regions (PRR) are required for optimal in vitro expression (Welter, J, F,, Crish, J. F., Agarwal, C., and Eckert, R. L, (1995) J. Biol. Chem. 270, 12614-12622; and Banks, E. B., Crish, J. F., Welter, J. F,, and Eckert, R. L. (1998) Biochem. J. 331, 61-68). We now present the complete sequence of these regions and evaluate their ability to drive in vivo transcription. Transgenes containing 5000 or 2473 base pairs of upstream regulatory region drive tissue- and differentiation-appropriate expression in stratifying surface epithelia, In contrast, transgenes containing 1953, 1333, 986, or 41 base pairs of upstream regulatory region are not expressed in surface epithelia, indicating that loss of the DRR (nucleotides -2474/-1953) results in loss of expression. Fusing the isolated DRR region directly to the hINV minimal promoter restores surface epithelial expression. Sequences downstream of the transcribed gene are not required for appropriate expression. The -1953/-41 segment influences the pattern of differentiation-dependent expression. The -986/-41 region, which includes the PRR, drives expression in internal epithelia.