Inhibition of the transcription factor Yin Yang 1 activity by S-nitrosation
Inhibition of the transcription factor Yin Yang 1 activity by S-nitrosation
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DOI：
10.1016/j.bbrc.2005.08.150
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发表时间：
2005-10-21
影响因子：
3.1
通讯作者：
Bonavida, B
中科院分区：
文献类型：
--
作者：
Hongo, F;Garban, H;Bonavida, B
用一氧化氮供体DETA/NONOate处理前列腺癌(CAP)细胞系PC-3、CL-1和DU-145后，细胞Fas表达上调，并对Fas配体CH-I激动剂单抗诱导的细胞凋亡敏感。先前的发现表明，转录抑制因子阴阳1(YY1)被NO抑制，负调控Fas转录[H.J.Garban，B.Bonavida，NO抑制转录抑制因子Fas启动子沉默区域的阴阳1结合活性：一氧化氮在人类肿瘤细胞Fas基因表达上调中起关键作用，J.167(2001)75-81]。YY1是一种锌指蛋白，因此我们推测NO通过与锌离子协调的关键半胱氨酸残基的S亚硝化来抑制YY1的活性。用EMSA法检测，DETA/NONOate处理PC-3细胞可抑制YY1的结构性DNA结合活性。此外，DETA/NONOate处理导致两种不同方法检测到的YY1的S亚硝化。基于DAN的方法检测了用抗YY1特异性抗体免疫沉淀的NO处理的肿瘤来源细胞裂解物，并用荧光法定量测定了释放的NO。第二种方法是用抗SNO半胱氨酸抗体免疫沉淀肿瘤细胞裂解物，免疫共沉淀物用抗YY1抗体免疫印迹。两种方法均显示DETA/NONOate对YY1细胞的S亚硝化作用显著高于对照细胞。用双色免疫荧光免疫组织化学方法进一步证实了YY1的S亚硝化反应。通过将siRNA YY1导入细胞，证实了YY1在Fas表达负调控中的直接作用。在没有DETA/NONOate处理的情况下，转基因细胞的Fas表达上调，并对CH-11诱导的细胞凋亡敏感。综上所述，这些结果表明，NO通过S亚硝化抑制YY1DNA结合活性，从而导致Fas表达上调，并使肿瘤细胞对Fas诱导的细胞凋亡增敏。(C)2005 Elsevier Inc.保留所有权利。
Treatment of several prostate cancer (CaP) cell lines (PC-3, CL-1, and DU-145) with the nitric oxide (NO) donor DETA/NONOate upregulated Fas expression and sensitized the CaP cells to the Fas ligand CH-I I agonist monoclonal antibody-induced apoptosis. Previous findings demonstrated that the transcription repressor Yin Yang 1 (YY1), which is inhibited by NO, negatively regulates Fas transcription [H.J. Garban, B. Bonavida, Nitric oxide inhibits the transcription repressor Yin-Yang 1 binding activity at the silencer region of the Fas promoter: a pivotal role for nitric oxide in the upregulation of Fas gene expression in human tumor cells, J. Immunol. 167 (2001) 75-81]. YY1 is a zinc finger protein and thus, we hypothesized that NO inhibits YY1 activity via S-nitrosation of critical cysteines residues coordinated by Zn2+. Treatment of PC-3 cells with DETA/NONOate inhibited the constitutive DNA-binding activity of YY1 as assessed by EMSA. Further, treatment with DETA/NONOate resulted in S-nitrosation of YY1 as detected by two different methods. The DAN-based method examined NO-treated tumor-derived cell lysates that were immunoprecipitated with an anti-YY1 specific antibody and the NO released was determined quantitatively by fluorometry. The second method consisted of immunoprecipitation of the tumor cell lysates by an anti-SNO cysteine antibody and the immunoprecipitate was immunoblotted with anti-YY1 antibody. Both methods revealed significant S-nitrosation of YY1 by DETA/NONOate treatment over control untreated cells. The S-nitrosation of YY1 was further corroborated by immunohistochemistry using dual color immunofluorescence. The direct role of YY1 in the negative regulation of Fas expression was demonstrated by transfection of cells with siRNA YY1. The transfectants exhibited upregulation of Fas expression in the absence of treatment with DETA/NONOate and were sensitized to CH-11-induced apoptosis. Altogether, these findings reveal that NO inhibits YY1 DNA-binding activity through S-nitrosation and consequently results in upregulation of Fas expression and tumor cell sensitization to Fas-induced apoptosis. (c) 2005 Elsevier Inc. All rights reserved.