title: Specification of Vδ and Vα usage by Tcra/Tcrd locus V gene segment promoters.
abstract: The Tcra/Tcrd locus undergoes V-Dδ-Jδ rearrangement in CD4(-)CD8(-) thymocytes to form the TCRδ chain of the γδ TCR and V-Jα rearrangement in CD4(+)CD8(+) thymocytes to form the TCRα-chain of the αβ TCR. Most V segments in the locus participate in V-Jα rearrangement, but only a small and partially overlapping subset participates in V-Dδ-Jδ rearrangement. What specifies any particular Tcra/Tcrd locus V gene segment as a Vδ, a Vα, or both is currently unknown. We tested the hypothesis that V segment usage is specified by V segment promoter-dependent chromatin accessibility in developing thymocytes. TRAV15/DV6 family V gene segments contribute to both the Tcrd and the Tcra repertoires, whereas TRAV12 family V gene segments contribute almost exclusively to the Tcra repertoire. To understand whether the TRAV15/DV6 promoter region specifies TRAV15/DV6 as a Vδ, we used gene targeting to replace the promoter region of a TRAV12 family member with one from a TRAV15/DV6 family member. The TRAV15/DV6 promoter region conferred increased germline transcription and histone modifications to TRAV12 in double-negative thymocytes and caused a substantial increase in usage of TRAV12 in Tcrd recombination events. Our results demonstrate that usage of TRAV15/DV6 family V gene segments for Tcrd recombination in double-negative thymocytes is regulated, at least in part, by intrinsic features of TRAV15/DV6 promoters, and argue that Tcra/Tcrd locus Vδ gene segments are defined by their local chromatin accessibility in CD4(-)CD8(-) thymocytes.
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710; and.
authors.forename: Abani Kanta
authors.initials: AK
authors.lastname: Naik
authors.affiliation: Department of Genetics, King Faisal Specialist Hospital & Research Centre, Riyadh 11211, Saudi Arabia krang001@mc.duke.edu ahawwari@kfshrc.edu.sa.
authors.forename: Abbas
authors.initials: A
authors.lastname: Hawwari
authors.affiliation: Department of Immunology, Duke University Medical Center, Durham, NC 27710; and krang001@mc.duke.edu ahawwari@kfshrc.edu.sa.
authors.forename: Michael S
authors.identifier: http://orcid.org/0000-0003-1292-254X
authors.initials: MS
authors.lastname: Krangel
mesh_terms: D000818:Animals; D015332:Gene Rearrangement, alpha-Chain T-Cell Antigen Receptor; D016694:Gene Rearrangement, delta-Chain T-Cell Antigen Receptor; D018390:Gene Targeting; D019673:Genes, T-Cell Receptor alpha; D019676:Genes, T-Cell Receptor delta; D056426:Genetic Loci; D051379:Mice; D011401:Promoter Regions, Genetic; D060168:Thymocytes / Q000166:cytology