Initiation of allelic exclusion by stochastic interaction of Tcrb alleles with repressive nuclear compartments

Abstract

The influence of repressive nuclear compartments on Tcrb rearrangement remains uncertain. Krangel and colleagues show that frequent stochastic, rather than directed, association of Tcrb alleles with repressive compartments promotes monoallelic recombination at this locus. Studies of antigen-receptor loci have linked directed monoallelic association with pericentromeric heterochromatin to the initiation or maintenance of allelic exclusion. Here we provide evidence for a fundamentally different basis for T cell antigen receptor-β (Tcrb) allelic exclusion. Using three-dimensional immunofluorescence in situ hybridization, we found that germline Tcrb alleles associated stochastically and at high frequency with the nuclear lamina or with pericentromeric heterochromatin in developing thymocytes and that such interactions inhibited variable–to–diversity-joining (Vβ-to-DβJβ) recombination before β-selection. The introduction of an ectopic enhancer into Tcrb resulted in fewer such interactions and impaired allelic exclusion. We propose that initial Vβ-to-DβJβ recombination events are generally monoallelic in developing thymocytes because of frequent stochastic, rather than directed, interactions of Tcrb alleles with repressive nuclear compartments. Such interactions may be essential for Tcrb allelic exclusion.