title: An anti-silencer- and SATB1-dependent chromatin hub regulates Rag1 and Rag2 gene expression during thymocyte development.
abstract: Rag1 and Rag2 gene expression in CD4(+)CD8(+) double-positive (DP) thymocytes depends on the activity of a distant anti-silencer element (ASE) that counteracts the activity of an intergenic silencer. However, the mechanistic basis for ASE activity is unknown. Here, we show that the ASE physically interacts with the distant Rag1 and Rag2 gene promoters in DP thymocytes, bringing the two promoters together to form an active chromatin hub. Moreover, we show that the ASE functions as a classical enhancer that can potently activate these promoters in the absence of the silencer or other locus elements. In thymocytes lacking the chromatin organizer SATB1, we identified a partial defect in Tcra gene rearrangement that was associated with reduced expression of Rag1 and Rag2 at the DP stage. SATB1 binds to the ASE and Rag promoters, facilitating inclusion of Rag2 in the chromatin hub and the loading of RNA polymerase II to both the Rag1 and Rag2 promoters. Our results provide a novel framework for understanding ASE function and demonstrate a novel role for SATB1 as a regulator of Rag locus organization and gene expression in DP thymocytes.
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710.
authors.forename: Bingtao
authors.initials: B
authors.lastname: Hao
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710.
authors.forename: Abani Kanta
authors.initials: AK
authors.lastname: Naik
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710.
authors.forename: Akiko
authors.initials: A
authors.lastname: Watanabe
authors.affiliation: RIKEN Centre for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710.
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, CA 94720.
authors.forename: Hunter W
authors.initials: HW
authors.lastname: Richards
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710.
authors.forename: Motonari
authors.initials: M
authors.lastname: Kondo
authors.affiliation: RIKEN Centre for Integrative Medical Sciences, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Taniuchi
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, CA 94720.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Kohwi
authors.affiliation: Life Sciences Division, Lawrence Berkeley National Laboratory, University of California, Berkeley, Berkeley, CA 94720.
authors.forename: Terumi
authors.initials: T
authors.lastname: Kohwi-Shigematsu
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710 krang001@mc.duke.edu.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Krangel
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology*; D002843:Chromatin / Q000235:genetics / Q000276:immunology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology*; D005786:Gene Expression Regulation / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics / Q000276:immunology*; D036961:Matrix Attachment Region Binding Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D018345:Mice, Knockout; D020218:Response Elements / Q000276:immunology*; D060168:Thymocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D036961:Matrix Attachment Region Binding Proteins; C499308:Rag2 protein, mouse; C469541:Satb1 protein, mouse; C064658:RAG-1 protein
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