Cutting Edge: Targeting of Vβ to Dβ Rearrangement by RSSs Can Be Mediated by the V(D)J Recombinase in the Absence of Additional Lymphoid-Specific Factors

Tillman, Robert E.; Wooley, Andrea L.; Khor, Bernard; Wehrly, Tara D.; Little, C.A.; Sleckman, Barry P.

doi:10.4049/jimmunol.170.1.5

Cited by 35 publications

(33 citation statements)

References 30 publications

(27 reference statements)

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“…in accordance with recent reports that were published while this manuscript was in preparation (23,24).…”

Section: Restricted V-d␤ Rearrangement Can Be Reproduced In Nonlymphosupporting

confidence: 79%

“…We cannot rule out that in vivo, additional factors participate in enforcing selective D gene usage, but our results suggest that the role of these factors, if any, is relatively minor compared with the effect of the Rag proteins. While this manuscript was in preparation, two groups have reported data that support the aforementioned conclusion (23,24). However, we have provided a series of additional data that go beyond the recently published reports.…”

Section: Discussionmentioning

confidence: 43%

“…However, recent elegant experiments with transgenic minilocus and knock-in mutants have successfully identified the flanking RS as the major determinants of restricted V-D rearrangement in the murine TCR␤ locus (11,22). In addition, while this manuscript was in preparation molecular studies have demonstrated that the Rag proteins are sufficient to reproduce these restrictions in ex vivo transfection and in vitro DNA cleavage assays (23,24).…”

Section: And References Therein)mentioning

confidence: 99%

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DNA-Rag Protein Interactions in the Control of Selective D Gene Utilization in the TCRβ Locus

Olaru

Patterson

Villey

et al. 2003

The Journal of Immunology

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Ordered assembly of Ag receptor genes by VDJ recombination is a key determinant of successful lymphocyte differentiation and function. Control of gene rearrangement has been traditionally viewed as a result of complex reorganization of the nucleochromatin mediated by several nuclear factors. Selective recombination of the variable (V) genes to the diversity (D), but not joining (J), gene segments within the TCRβ locus has been shown to be controlled by recombination signal (RS) sequences that flank the gene segments. Through ex vivo and in vitro recombination assays, we demonstrate that the Rag proteins can discriminate between the RS of the D and J genes and enforce selective D gene incorporation into the TCRβ variable domain in the absence of other nuclear factors or chromatin structure. DNA binding studies indicate that discrimination is not simply caused by higher affinity binding of the Rag proteins to the isolated 12RS of the D as opposed to the J genes. Furthermore, we also demonstrate that the 12RS within the TCRβ locus is functionally inferior to the consensus 12RS. We propose that selective gene segment usage is controlled at the level of differential assembly and/or stability of synaptic RS complexes, and that evolutionary “deterioration” of the RS motifs may have been important to allow the VDJ recombinase to exert autonomous control over gene segment use during gene rearrangement.

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“…in accordance with recent reports that were published while this manuscript was in preparation (23,24).…”

Section: Restricted V-d␤ Rearrangement Can Be Reproduced In Nonlymphosupporting

confidence: 79%

Section: Discussionmentioning

confidence: 43%

Section: And References Therein)mentioning

confidence: 99%

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DNA-Rag Protein Interactions in the Control of Selective D Gene Utilization in the TCRβ Locus

Olaru

Patterson

Villey

et al. 2003

The Journal of Immunology

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“…Surprisingly, however, our transfection experiments demonstrate that the Rag proteins, by themselves, are insufficient to enforce TCR␦ D gene incorporation. This observation stands in striking contrast to the TCR␤ locus where isolated RS/coding end combinations and their interactions with the Rag proteins were sufficient to reproduce the specific patterns of both V-D and D-J recombination (12)(13)(14)26).…”

Section: Discussioncontrasting

confidence: 42%

Beyond the 12/23 Rule of VDJ Recombination Independent of the Rag Proteins

Olaru

Petrie²,

Livák³

2005

The Journal of Immunology

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The combinatorial repertoire of AgRs is established through somatic recombination of V, D, and J gene segments during lymphocyte development. Incorporation of D segments into IgH, TCRβ, and TCRδ chains also contributes to junctional diversification by substantially extending the length of the third CDR. The V, D, and J gene segments are flanked by recombination signals (RS) of 12- or 23-mer spacer length that direct recombination according to the 12/23 rule. D genes in the TCRβ and TCRδ loci are flanked by a 12RS and 23RS, and their incorporation is controlled by mechanisms “beyond the 12/23 rule.” In the TCRβ locus, selective interactions between Rag proteins and the RS flanking the V-D and D-J genes, respectively, are sufficient to enforce D gene usage. In this article, we report that in the TCRδ locus, the Rag proteins are not the major determinant of D gene incorporation. In developing mouse and human thymocytes, the two Dδ genes rearrange predominantly to form D-D coding joints. In contrast, when tested in ex vivo transfection assays in a nonlymphoid cell line, the flanking RS mediate deletion, rather than incorporation, of the two D genes on both exogenous recombination substrates and the endogenous locus. These results suggest that selective Rag-RS interactions are not the sole regulators of D gene segment incorporation, and additional, perhaps lymphocyte-specific, mechanisms exist that allow proper shaping of the primary AgR repertoire.

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“…V(D)J recombination is initiated by the recombinase activating proteins encoded by the recombinase activating genes, Rag-1 and Rag-2 which are only expressed in cells undergoing antigenic receptor rearrangement [22][23][24]. Rag proteins bind to recombination signal sequences (RSSs) that flank the V, D and J gene segments and initiate double-strand breaks (DSB) [25].…”

mentioning

confidence: 99%

MYB IS Required for B-Selection During Thymopoises

Duley¹

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Abstractc-Myb is a transcription factor required during hematopoiesis and is crucial for multiple stages of thymopoiesis. Understanding c-Myb function during hematopoiesis has been greatly impeded due to the lack of a tractable genetic system. We have used the Cre/loxP approach to conditional mutagenesis to study c-Myb function during thymopoiesis. Thymocyte specific inactivation of the Myb locus demonstrated a block in DN3 to DN4 differentiation concomitant with impaired Vβ to DJβ recombination of the Tcrb locus suggesting that the inability to generate a TCRβ protein blocked DN3 differentiation. However, some DN3 thymocytes

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Cutting Edge: Targeting of Vβ to Dβ Rearrangement by RSSs Can Be Mediated by the V(D)J Recombinase in the Absence of Additional Lymphoid-Specific Factors

Cited by 35 publications

References 30 publications

DNA-Rag Protein Interactions in the Control of Selective D Gene Utilization in the TCRβ Locus

DNA-Rag Protein Interactions in the Control of Selective D Gene Utilization in the TCRβ Locus

Beyond the 12/23 Rule of VDJ Recombination Independent of the Rag Proteins

MYB IS Required for B-Selection During Thymopoises

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