Petricoin E, David M, Igarashi K, Benjamin C, Ling L, Goelz S, Finbloom D S, Larner A C
Division of Cytokine Biology, Center for Biologics Evaluation and Research, Bethesda, Maryland 20892, USA.
Mol Cell Biol. 1996 Apr;16(4):1419-24. doi: 10.1128/MCB.16.4.1419.
Previous studies have indicated that the expression of viral oncoproteins, cell transformation, or phorbol ester treatment of cells can inhibit alpha/beta interferon (IFN-alpha/beta)-induced gene expression. The mechanisms by which these promoters of cell growth exert their inhibitory effects vary, but in most instances they involve a disruption of the IFN-alpha/beta-induced transcription complex ISGF3 such that the DNA-binding component of this complex (the 48-kDa ISGF3gamma protein) does not bind to the interferon-stimulated response element (ISRE). In this report, we demonstrated that phorbol ester treatment of human peripheral blood monocytes dramatically inhibits activation of IFN-alpha/B-stimulated early response genes but by a mechanism which does not involve abrogation of the ISRE binding of ISGF3gamma. Phorbol ester treatment of monocytes inhibited IFN alpha-stimulated tyrosine phosphorylation of the transcription factors Stat1alpha, Stat2, and Stat3 and of the tyrosine kinase Tyk2 but had no effect on IFN-gamma activation of Stat1alpha. IFNalpha-stimulated tyrosine phosphorylation of Jak1 and the alpha subunit of the IFN-alpha receptor were unaffected by phorbol 12-myristate 13-acetate (PMA). Moreover, PMA caused the dephosphorylation of Tyk2 but not of Jak1, which was activated by IFN. Pretreatment of cells with vanadate prevented the effects of PMA with regard to PMA-induced Tyk2 dephosphorylation. These observations suggest that PMA exerts its inhibitory effects by activation of a tyrosine phosphatase which selectively regulates Tyk2 but not Jak1 activity.
先前的研究表明,病毒癌蛋白的表达、细胞转化或佛波酯处理细胞可抑制α/β干扰素(IFN-α/β)诱导的基因表达。这些细胞生长促进剂发挥抑制作用的机制各不相同,但在大多数情况下,它们涉及破坏IFN-α/β诱导的转录复合物ISGF3,使得该复合物的DNA结合成分(48 kDa的ISGF3γ蛋白)不与干扰素刺激反应元件(ISRE)结合。在本报告中,我们证明佛波酯处理人外周血单核细胞可显著抑制IFN-α/B刺激的早期反应基因的激活,但其机制并不涉及废除ISGF3γ与ISRE的结合。佛波酯处理单核细胞可抑制IFNα刺激的转录因子Stat1α、Stat2和Stat3以及酪氨酸激酶Tyk2的酪氨酸磷酸化,但对IFN-γ激活Stat1α没有影响。IFNα刺激的Jak1和IFN-α受体α亚基的酪氨酸磷酸化不受佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)的影响。此外,PMA导致Tyk2去磷酸化,但不导致IFN激活的Jak1去磷酸化。用钒酸盐预处理细胞可防止PMA对PMA诱导的Tyk2去磷酸化的影响。这些观察结果表明,PMA通过激活一种酪氨酸磷酸酶发挥其抑制作用,该酪氨酸磷酸酶选择性调节Tyk2的活性而不调节Jak1的活性。