Coding sequence composition flanking either signal element alters V(D)J recombination efficiency.
Lymphoid V(D)J rearrangement is targeted by recombination signal sequences (RSS) bordering V, D or J exons. We demonstrate that the DNA composition of flanking coding positions, particularly poly(A) or poly(T) stretches at one or both RSS, diminishes V(D)J recombination up to 100-fold. Positionally correct cleavages occur in the inhibited reactions, since the junctions formed show the same frequency of precision as uninhibited reactions. Open/shut cleavage/rejoining is not increased at a normal RSS in substrates containing inhibitory A/T homopolymers versus random sequence at a second RSS. Thus recombinase action at both cleavage sites is severely disrupted by modified coding sequences.
Boubnov
N V
NV
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
Wills
Z P
ZP
Weaver
D T
DT
D001402:B-Lymphocytes / Q000201:enzymology; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D015322:Gene Rearrangement, B-Lymphocyte / Q000235:genetics*; D019426:Integrases; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D045522:Recombinases; D011995:Recombination, Genetic / Q000235:genetics*; D013698:Templates, Genetic; D014407:Tumor Cells, Cultured
D009838:Oligodeoxyribonucleotides; D045522:Recombinases; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C115006:integron integrase IntI1