Productive coupling of accessible Vbeta14 segments and DJbeta complexes determines the frequency of Vbeta14 rearrangement.
To elucidate mechanisms that regulate Vbeta rearrangement, we generated and analyzed mice with a V(D)J recombination reporter cassette of germline Dbeta-Jbeta segments inserted into the endogenous Vbeta14 locus (Vbeta14(Rep)). As a control, we first generated and analyzed mice with the same Dbeta-Jbeta cassette targeted into the generally expressed c-myc locus (c-myc(Rep)). Substantial c-myc(Rep) recombination occurred in both T and B cells and initiated concurrently with endogenous Dbeta to Jbeta rearrangements in thymocytes. In contrast, Vbeta14(Rep) recombination was restricted to T cells and initiated after endogenous Dbeta to Jbeta rearrangements, but concurrently with endogenous Vbeta14 rearrangements. Thus, the local chromatin environment imparts lineage and developmental stage-specific accessibility upon the inserted reporter. Although Vbeta14 rearrangements occur on only 5% of endogenous TCRbeta alleles, the Vbeta14(Rep) cassette underwent rearrangement on 80-90% of alleles, supporting the suggestion that productive coupling of accessible Vbeta14 segments and DJbeta complexes influence the frequency of Vbeta14 rearrangements. Strikingly, Vbeta14(Rep) recombination also occurs on TCRbeta alleles lacking endogenous Vbeta to DJbeta rearrangements, indicating that Vbeta14 accessibility per se is not subject to allelic exclusion.
Ranganath
Sheila
S
Howard Hughes Medical Institute, The Children's Hospital, CBR Institute for Biomedical Research, and Department of Genetics, Harvard University Medical School, Boston, MA 02115, USA.
Carpenter
Andrea C
AC
Gleason
Megan
M
Shaw
Albert C
AC
Bassing
Craig H
CH
Alt
Frederick W
FW
D000818:Animals; D002478:Cells, Cultured; D002875:Chromosomes / Q000235:genetics; D022801:Complementarity Determining Regions / Q000235:genetics; D053595:Embryonic Stem Cells / Q000378:metabolism; D015322:Gene Rearrangement, B-Lymphocyte; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor / Q000235:genetics*; D017930:Genes, Reporter; D007133:Immunoglobulin Joining Region / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D016254:Mutagenesis, Insertional; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism*; D011995:Recombination, Genetic; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism
D022801:Complementarity Determining Regions; D007133:Immunoglobulin Joining Region; D007135:Immunoglobulin Variable Region; D010446:Peptide Fragments