人RAG2基因5′近端染色质结构定量差异分析及可接近区域转录调控机制研究

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Abstract Objective To study molecular mechanism of the regulation of RAG2 expression regulated the human recombination activating gene 2(RAG2)5′proximal chromatin accessible areas.Methods To compare the open states change of chromatin of RAG positive and negative lymphocytes as well as non lymphocyte chromatin,chromatin accessibility by real-time polymerase chain reaction(CHART-PCR)was used for DNaseⅠanalysis of RAG2 gene 5′proximal regions of hypersensitive sites(DHS).Double luciferases reporter assay was used to examine the transcriptional regulation effects of the DHS regions.Electrophoresis mobility shift assay and chromatin immunoprecipitation assays were used to detect the binding of GATA3 in vitro or in vivo.PCR site-directed mutant assay and dominant-negative mutant assay were used to identify the regulatory effects of GATA3 on the reporter gene(in vitro),as well as on RAG2 gene(in vivo). Results In T and B cells,the chromatin structures of 5′proximal region of human RAG2 were in distinct open states.In RAG positive T cells,the activity of RAG2 core promoter corresponded to its specific DHS.GATA3,a T-cell specific transfactor,bound to a highly conserved sequence within the specific DHS,and increased the expression of the reporter genes as well as RAG2 mRNA of the T cells in vivo.Conclusion RAG2 gene 5′proximal area through open states change of chromatin regulate the specificity of the gene expression,GATA3 regulates the expression of endogenous RAG2 in T cells by binding to highly conserved DHS regions.

Key words： Recombination activating gene Chromatin accessibility Promoter Cis-acting element Transcription factor T lymphocyte

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	XUE Wen-yu
	ZENG Yan
	WEI Xing-cheng

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XUE Wen-yu,ZENG Yan,WEI Xing-cheng. Quantitative difference analysis of chromatin structure across the 5′proximal region of human RAG2 and study of its transcriptional regulatory mechanisms on the accessible areas[J]. 中国当代医药, 2017, 24(3): 7-12转16.

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http://dangdaiyiyao.com/EN/ OR http://dangdaiyiyao.com/EN/Y2017/V24/I3/7

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