Regulation of human involucrin promoter activity by novel protein kinase C isoforms
Regulation of human involucrin promoter activity by novel protein kinase C isoforms
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DOI：
10.1074/jbc.275.3.1601
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发表时间：
2000-01-21
影响因子：
4.8
通讯作者：
Eckert, RL
中科院分区：
文献类型：
--
作者：
Efimova, T;Eckert, RL
当用分化剂12-O-十四烷酰基佛波醇-13-乙酸酯（TPA）处理角质形成细胞时，人外皮蛋白（hINV）mRNA水平和启动子活性增加。该应答通过靶向激活蛋白1的p38促分裂原激活蛋白激酶依赖性途径介导（Efimova，T.，LaCelle，P. T.，韦尔特，J.F.，和Eckert，R. L.（1998）J.Biol.Chem.273，24387-24395），在本研究中，我们检查了各种PKC同种型在该调节中的作用。转染编码新型PKC亚型δ、β和η的表达质粒增加hINV启动子活性。相比之下，无论是传统的PKC亚型（α，β和γ），也不是非典型亚型（ζ）调节启动子活性。与这些观察结果一致，启动子活性被PKC δ选择性抑制剂rottlerin抑制，但不被Go-6976抑制，Go-6976是一种常规PRC亚型的抑制剂，并且新型PKC亚型依赖性启动子激活被显性负性PKC δ抑制。这种调节似乎是生理学上重要的，因为用PKC δ、PKC β或PKC β转染角质形成细胞增加内源性hINV基因的表达。当用TPA处理PKC β-或-β-转染的细胞时，观察到协同启动子活化（大于或等于100倍）。相比之下，PKC δ依赖性反应更复杂，因为根据PKC δ浓度观察到激活或抑制。
Human involucrin (hINV) mRNA level and promoter activity increase when keratinocytes are treated with the differentiating agent, 12-O-tetradecanoylphorbol-13-acetate (TPA). This response is mediated via a p38 mitogen-activated protein kinase-dependent pathway that targets activator protein 1 (Efimova, T., LaCelle, P. T., Welter, J. F., and Eckert, R. L. (1998) J. Biol, Chem. 273, 24387-24395), In the present study we examine the role of various PKC isoforms in this regulation. Transfection of expression plasmids encoding the novel PKC isoforms delta, epsilon, and eta increase hINV promoter activity. In contrast, neither conventional PKC isoforms (alpha, beta, and gamma) nor the atypical isoform (zeta) regulate promoter activity. Consistent with these observations, promoter activity is inhibited by the PKC delta-selective inhibitor, rottlerin, but not by Go-6976, an inhibitor of conventional PRC isoforms, and novel PKC isoform-dependent promoter activation is inhibited by dominant-negative PKC delta. This regulation appears to be physiologically important, as transfection of keratinocytes with PKC delta, -epsilon, or -eta increases expression of the endogenous hINV gene. Synergistic promoter activation (greater than or equal to 100-fold) is observed when PKC epsilon- or -eta-transfected cells are treated with TPA. In contrast, the PKC delta-dependent response is more complex as either activation or inhibition is observed, depending upon PKC delta concentration.