Benton K D, Hermann R J, Vomhof-DeKrey E E, Haring J S, Van der Steen T, Smith J, Dovat S, Dorsam G P
Department of Chemistry and Molecular Biology, North Dakota State University, Fargo, ND 58108, USA.
Regul Pept. 2009 Nov 27;158(1-3):68-76. doi: 10.1016/j.regpep.2009.08.010. Epub 2009 Sep 1.
T cells express receptors for neuropeptides that mediate immunological activities. Vasoactive intestinal peptide receptor-1 (VPAC1), the prototypical group II G protein coupled receptor, binds two neuropeptides with high-affinity, called vasoactive intestinal peptide and pituitary adenylate cyclase activating polypeptide. During T cell signaling, VPAC1 mRNA expression levels are significantly downregulated through a Src kinase dependent mechanism, thus altering the sensitivity for these neuropeptides during an immune reaction. Presently, it is unknown whether the mechanism that regulates VPAC1 during T cell signaling involves epigenetic changes. Therefore, we hypothesized that the epigenetic landscape consisting of diacetylation at H3K9/14 and trimethylation at H3K4, two transcriptionally permissive histone modifications, would parallel VPAC1 expression showing high enrichment in untreated T cells, but lower enrichment in alpha-CD3 treated T cells. To this end, quantitative chromatin immunoprecipitation (ChIP) analysis of H3K9/14ac and H3K4me3 was conducted using purified CD4(+) T cells, with CD45R(+) B cells as a negative control. Our data revealed that these histone modifications at the VPAC1 promoter did indeed parallel its mRNA levels between T and B lymphocytes, but did not decrease during T cell signaling. Collectively, these data strongly imply a euchromatin nuclear position for the VPAC1 locus irrespective of the activation status of T cells.
T细胞表达介导免疫活性的神经肽受体。血管活性肠肽受体-1(VPAC1)是典型的II类G蛋白偶联受体,能高亲和力结合两种神经肽,即血管活性肠肽和垂体腺苷酸环化酶激活多肽。在T细胞信号传导过程中,VPAC1 mRNA表达水平通过Src激酶依赖机制显著下调,从而在免疫反应期间改变对这些神经肽的敏感性。目前,尚不清楚T细胞信号传导过程中调节VPAC1的机制是否涉及表观遗传变化。因此,我们推测由H3K9/14二乙酰化和H3K4三甲基化组成的表观遗传格局,这两种转录允许的组蛋白修饰,将与VPAC1表达平行,在未处理的T细胞中显示高富集,但在α-CD3处理的T细胞中富集较低。为此,使用纯化的CD4(+) T细胞进行H3K9/14ac和H3K4me3的定量染色质免疫沉淀(ChIP)分析,以CD45R(+) B细胞作为阴性对照。我们的数据显示,VPAC1启动子处的这些组蛋白修饰确实在T淋巴细胞和B淋巴细胞之间与其mRNA水平平行,但在T细胞信号传导过程中并未降低。总体而言,这些数据强烈暗示VPAC1基因座处于常染色质核位置,与T细胞的激活状态无关。