T cell receptor DJ but not VDJ rearrangement within a recombination substrate introduced into a pre-B cell line.
To elucidate mechanisms that regulate ordered and tissue-specific assembly of Ig and TCR variable region gene segments, we have introduced a recombination substrate comprised of germline TCR beta V, D, and J gene segments into an Abelson murine leukemia virus-transformed pre-B cell line that actively rearranges endogenous Ig H chain variable region gene segments but does not rearrange endogenous light chain or TCR variable region gene segments. We find that these cells efficiently join D beta segments to J beta segments within the mini-locus, but that they do not make any detectable site-specific rearrangements of the introduced V beta segment even though it is closely linked in the same construct to the D beta. These findings suggest that factors necessary for V beta to (D beta)J beta joining may be absent in these pre-B cells and also imply that the order in which TCR V beta, D beta, and J beta segments are rearranged can be influenced by factors other than the 12/23 recombination rule. Furthermore, in agreement with the an accessibility model of VDJ recombinase control, the D beta region of the construct was found to be relatively more sensitive to DNAase I digestion in isolated nuclei when compared to the unrearranged V beta region.
Ferrier
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Howard Hughes Medical Institute, New York, New York 10032.
Covey
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LR
Suh
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Winoto
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A
Hood
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Alt
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FW
D000011:Abelson murine leukemia virus; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D002472:Cell Transformation, Viral; D003851:Deoxyribonucleases; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D008957:Models, Genetic; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013937:Thymidine Kinase / Q000235:genetics
D013937:Thymidine Kinase; D003851:Deoxyribonucleases