CARM1 but not Its Enzymatic Activity Is Required for Transcriptional Coactivation of NF-κB-Dependent Gene Expression
CARM1 but not Its Enzymatic Activity Is Required for Transcriptional Coactivation of NF-κB-Dependent Gene Expression
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DOI：
10.1016/j.jmb.2009.09.032
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发表时间：
2009-12-04
影响因子：
5.6
通讯作者：
Hottiger, Michael O.
中科院分区：
文献类型：
--
作者：
Jayne, Sandrine;Rothgiesser, Karin M.;Hottiger, Michael O.
共激活剂相关精氨酸甲基转移酶 1 (CARM1) 属于蛋白质精氨酸甲基转移酶家族。据报道，它可以甲基化组蛋白和非组蛋白，因此参与转录激活和 mRNA 降解/稳定性。在这里，我们报告了 carm1-/- 细胞与野生型 CARM1 或 CARM1 酶促无活性突变体的遗传互补，以研究 CARM1 及其酶活性对核因子 kappa B (NF-kappa B) 依赖性基因表达的需求。使用定制微阵列和定量逆转录 PCR，我们可以定义需要 CARM1 才能正确表达的 NF-kappa B 靶基因子集。虽然一些肿瘤坏死因子-α-和佛波醇-12-肉豆蔻酸-13-乙酸酯/离子霉素诱导的 NF-κ B 靶基因是 CARM1 依赖性的，但 CARM1 酶活性对于基因表达来说是可有可无的。有趣的是，CARM1 并不是依赖刺激的 RelA/p65 向染色质的募集所必需的，这表明 CARM1 在蛋白质复合物的稳定方面发挥了相当大的作用。总之，我们的结果证实了 CARM1 作为转录辅助因子的重要性，但不涉及其催化活性。 (C) 2009 Elsevier Ltd. 保留所有权利。
Coactivator-associated arginine methyltransferase 1 (CARM1) belongs to the protein arginine methyltransferase family. It was reported to methylate histone as well as non-histone proteins and thus to be involved in transcriptional activation and mRNA degradation/stability. Here we report the genetic complementation of carm1-/- cells with wild-type CARM1 or an enzymatic inactive mutant of CARM1 to investigate the requirement of CARM1 and its enzymatic activity for nuclear factor kappa B (NF-kappa B)-dependent gene expression. Using custom microarray and quantitative reverse transcription PCR, we could define a subset of NF-kappa B target genes that required CARM1 for their proper expression. Although several tumor necrosis factor-alpha- and phorbol-12-myristate-13-acetate/ionomycin-induced NF-kappa B target genes are CARM1 dependent, CARM1 enzymatic activity was dispensable for gene expression. Interestingly, CARM1 was not required for the stimulus-dependent recruitment of RelA/p65 to chromatin, suggesting that CARM1 is rather contributing in protein complex stabilization. Together, our results confirm the importance of CARM1 as transcriptional cofactor without the involvement of its catalytic activity. (C) 2009 Elsevier Ltd. All rights reserved.