Meyer M, Caselmann W H, Schlüter V, Schreck R, Hofschneider P H, Baeuerle P A
Max-Planck-Institute for Biochemistry, Department of Virus Research, Martinsried, Germany.
EMBO J. 1992 Aug;11(8):2991-3001. doi: 10.1002/j.1460-2075.1992.tb05369.x.
C-terminal truncation of the middle surface antigen from hepatitis B virus (MHBs) gives rise to a novel transactivating protein, called MHBst. In this study we show that MHBst like the HBx protein of HBV, can cause nuclear appearance of NF-kappa B DNA binding activity and induce various kappa B-controlled reporter genes. While an inhibitor of protein kinase C could not block gene induction by MHBst, the antioxidants N-acetyl-L-cysteine (NAC) and pyrrolidine dithiocarbamate (PDTC) could potently suppress transactivation at mM and microM concentrations, respectively. Also, kappa B-dependent gene induction by the transactivator HBx was blocked. The effects were selective because PDTC did not interfere with MHBst and HBx-induced activation of the c-fos promoter/enhancer, nor with the basal activity of several other reporter genes lacking functional NF-kappa B binding motifs. Our data suggest that induction of a prooxidant state is crucial for the activation of NF-kappa B by MHBst and HBx and might be related to the hepatocarcinogenic potential of the viral proteins. MHBst had a subcellular localization unusual for a viral transactivator: it appeared to be an integral membrane protein of the endoplasmic reticulum.
乙型肝炎病毒(HBV)中表面抗原(MHBs)的C末端截短会产生一种名为MHBst的新型反式激活蛋白。在本研究中,我们发现MHBst与HBV的HBx蛋白一样,可导致NF-κB DNA结合活性出现于细胞核内,并诱导各种κB控制的报告基因。虽然蛋白激酶C抑制剂不能阻断MHBst对基因的诱导作用,但抗氧化剂N-乙酰-L-半胱氨酸(NAC)和吡咯烷二硫代氨基甲酸盐(PDTC)分别在毫摩尔和微摩尔浓度下能有效抑制反式激活。此外,反式激活剂HBx对κB依赖基因的诱导也被阻断。这些作用具有选择性,因为PDTC不干扰MHBst和HBx诱导的c-fos启动子/增强子的激活,也不影响其他几个缺乏功能性NF-κB结合基序的报告基因的基础活性。我们的数据表明,诱导促氧化状态对于MHBst和HBx激活NF-κB至关重要,并且可能与病毒蛋白的致癌潜力有关。MHBst具有一种对于病毒反式激活剂而言不寻常的亚细胞定位:它似乎是内质网的一种整合膜蛋白。