1. Hierarchical assembly and disassembly of a transcriptionally active RAG locus in CD4+CD8+ thymocytes
- Author
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Aaron Lucander, Abani Kanta Naik, Aaron Byrd, and Michael S. Krangel
- Subjects
0301 basic medicine ,Transcription, Genetic ,CD8 Antigens ,Immunology ,chemical and pharmacologic phenomena ,GATA3 Transcription Factor ,Response Elements ,Article ,Recombination-activating gene ,Ikaros Transcription Factor ,Mice ,03 medical and health sciences ,0302 clinical medicine ,RAG2 ,hemic and lymphatic diseases ,Gene expression ,Basic Helix-Loop-Helix Transcription Factors ,Animals ,Immunology and Allergy ,Research Articles ,Homeodomain Proteins ,Transcriptionally active chromatin ,Thymocytes ,Chemistry ,T-cell receptor ,hemic and immune systems ,Promoter ,3. Good health ,Chromatin ,Cell biology ,DNA-Binding Proteins ,030104 developmental biology ,Gene Expression Regulation ,Genetic Loci ,CD4 Antigens ,tissues ,CD8 ,030215 immunology - Abstract
Naik et al. show that GATA3, Runx1, and E2A are essential for hierarchical assembly of a transcriptionally active RAG locus chromatin hub in CD4+CD8+ thymocytes. Signal-dependent down-regulation of RAG expression is associated with hub disassembly and depends on Ikaros., Expression of Rag1 and Rag2 is tightly regulated in developing T cells to mediate TCR gene assembly. Here we have investigated the molecular mechanisms governing the assembly and disassembly of a transcriptionally active RAG locus chromatin hub in CD4+CD8+ thymocytes. Rag1 and Rag2 gene expression in CD4+CD8+ thymocytes depends on Rag1 and Rag2 promoter activation by a distant antisilencer element (ASE). We identify GATA3 and E2A as critical regulators of the ASE, and Runx1 and E2A as critical regulators of the Rag1 promoter. We reveal hierarchical assembly of a transcriptionally active chromatin hub containing the ASE and RAG promoters, with Rag2 recruitment and expression dependent on assembly of a functional ASE–Rag1 framework. Finally, we show that signal-dependent down-regulation of RAG gene expression in CD4+CD8+ thymocytes depends on Ikaros and occurs with disassembly of the RAG locus chromatin hub. Our results provide important new insights into the molecular mechanisms that orchestrate RAG gene expression in developing T cells.
- Published
- 2018