Cell type-specific chromatin structure determines the targeting of V(D)J recombinase activity in vitro.
A common V(D)J recombinase that recognizes a conserved recombination signal sequence (RSS) mediates the assembly of immunoglobulin (Ig) and T cell receptor (TCR) genes in B and T cell precursors. The rearrangement of particular Ig and TCR gene segments, however, is tightly regulated with respect to cell lineage and developmental stage. Using an in vitro system, we analyzed recombinase cleavage of RSSs flanking Ig and TCR gene segments in nuclei. We found that both the lineage-specificity and temporal ordering of gene rearrangement is reflected in the accessibility of RSSs within chromatin to in vitro cleavage.
Stanhope-Baker
P
P
Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Hudson
K M
KM
Shaffer
A L
AL
Constantinescu
A
A
Schlissel
M S
MS
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D002417:Cattle; D002457:Cell Extracts; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000235:genetics*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D015322:Gene Rearrangement, B-Lymphocyte / Q000235:genetics*; D015329:Gene Rearrangement, T-Lymphocyte / Q000235:genetics*; D005803:Genes, Immunoglobulin / Q000235:genetics; D018398:Homeodomain Proteins; D007144:Immunoglobulin J-Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011506:Proteins / Q000502:physiology; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D013950:Thymus Gland; D045567:VDJ Recombinases
D002457:Cell Extracts; D002843:Chromatin; D018398:Homeodomain Proteins; D007144:Immunoglobulin J-Chains; D007145:Immunoglobulin kappa-Chains; D007148:Immunoglobulin mu-Chains; D011506:Proteins; D011948:Receptors, Antigen, T-Cell; C064658:RAG-1 protein; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases