Kučerová-Levisohn Martina, Knirr Stefan, Mejia Rosa I, Ortiz Benjamin D
Department of Biological Sciences, City University of New York, Hunter College, New York, New York, United States of America; Doctoral Program in Biology, City University of New York, Graduate Center, New York, New York, United States of America.
Department of Biological Sciences, City University of New York, Hunter College, New York, New York, United States of America.
PLoS One. 2015 Jul 15;10(7):e0132856. doi: 10.1371/journal.pone.0132856. eCollection 2015.
Much progress has been made in understanding the important cis-mediated controls on mouse TCRα gene function, including identification of the Eα enhancer and TCRα locus control region (LCR). Nevertheless, previous data have suggested that other cis-regulatory elements may reside in the locus outside of the Eα/LCR. Based on prior findings, we hypothesized the existence of gene regulatory elements in a 3.9-kb region 5' of the Cα exons. Using DNase hypersensitivity assays and TCRα BAC reporter transgenes in mice, we detected gene regulatory activity within this 3.9-kb region. This region is active in both thymic and peripheral T cells, and selectively affects upstream, but not downstream, gene expression. Together, these data indicate the existence of a novel cis-acting regulatory complex that contributes to TCRα transgene expression in vivo. The active chromatin sites we discovered within this region would remain in the locus after TCRα gene rearrangement, and thus may contribute to endogenous TCRα gene activity, particularly in peripheral T cells, where the Eα element has been found to be inactive.
在理解对小鼠TCRα基因功能重要的顺式介导调控方面已取得了很大进展,包括鉴定Eα增强子和TCRα基因座控制区(LCR)。然而,先前的数据表明其他顺式调控元件可能存在于Eα/LCR之外的基因座中。基于先前的发现,我们推测在Cα外显子5'端的3.9 kb区域存在基因调控元件。利用小鼠中的DNase超敏反应分析和TCRαBAC报告转基因,我们在这个3.9 kb区域内检测到了基因调控活性。该区域在胸腺T细胞和外周T细胞中均有活性,并选择性地影响上游而非下游的基因表达。总之,这些数据表明存在一种新型的顺式作用调控复合体,它有助于体内TCRα转基因的表达。我们在该区域内发现的活性染色质位点在TCRα基因重排后仍会保留在基因座中,因此可能有助于内源性TCRα基因的活性,特别是在外周T细胞中,在那里已发现Eα元件无活性。