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E protein binding at the Tcra enhancer promotes Tcra repertoire diversity

ABSTRACT: E protein binding at the Tcra enhancer promotes Tcra repertoire diversity

PROVIDER: PRJNA943495 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23821624_1.fastq.gz	Fastqsanger.gz
	SRR23821624_2.fastq.gz	Fastqsanger.gz

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E protein binding at the Tcra enhancer promotes Tcra repertoire diversity

Project description:The Tcra gene enhancer (Eα) is required for Tcra rearrangment. Our study assesses the role of E protein binding at Eα. Tcra repertoire sequencing was used to determine whether loss of Eα E protein binding sites alters recombination at Tcra.

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E protein binding at the <i>Tcra</i> enhancer promotes <i>Tcra</i> repertoire diversity.

Project description:V(D)J recombination of antigen receptor loci is a highly developmentally regulated process. During T lymphocyte development, recombination of the Tcra gene occurs in CD4+CD8+ double positive (DP) thymocytes and requires the Tcra enhancer (Eα). E proteins are known regulators of DP thymocyte development and have three identified binding sites in Eα. To understand the contribution of E proteins to Eα function, mutants lacking one or two of the respective binding sites were generated. The double-binding site mutant displayed a partial block at the positive selection stage of αβ T cell development. Further investigation revealed loss of germline transcription within the Tcra locus at the Jα array, along with dysregulated primary and impaired secondary Vα-Jα rearrangement. Eα E protein binding increases Tcra locus accessibility and regulates TCRα recombination, thus directly promoting Tcra repertoire diversity.

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Tcrd rearrangement redirects a processive Tcra recombination program to expand the Tcra repertoire

Project description:The Tcra-Tcrd locus undergoes Tcrd rearrangement in DN thymocytes followed by primary and secondary Tcra rearrangements in DP thymocytes. Here we reveal the mechanisms underpinning combinatorial diversity in the Tcra repertoire. We show that V-alpha and J-alpha segments on individual Tcra alleles are used in stepwise and coordinated proximal-to-distal progressions during primary and secondary rearrangements, substantially constraining combinatorial diversity. Notably, Tcrd recombination in DN thymocytes diversifies the Tcra repertoire by truncating the V-alpha array to permit otherwise disfavored V-J combinations. Such diversification is important, because Trav1-Traj33+ mucosa-associated invariant T cells are depleted when Tcrd rearrangement is impaired. Our results reveal an important biological advantage conferred by the nested organization of Tcrd and Tcra gene segments.

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A role of the CTCF binding site at enhancer Ea in Tcra rearrangement

Project description:We found that the CTCF binding site (named as EACBE) at the enhancer Eα of Tcra is essential for normal Tcra rearrangement.The EACBE deletion impaired the TAD structure of the 3’ region of the Tcra/d locus and the interactions between the proximal Vα genes and the Jα genes without changing the accessibility of the Jα array.

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Trav15-dv6 family Tcrd rearrangements diversify the Tcra repertoire

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ROR t directs upregulation of RAG gene expression in DP thymocytes and serves as a multi-faceted regulator of the Tcra repertoire.

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