Higher-order chromatin structure-dependent repair of DNA double-strand breaks: involvement of the V(D)J recombination double-strand break repair pathway.

Repair of DNA double-strand breaks (DSBs) is linked to the V(D)J recombination pathway through investigations of radiation-sensitive mutants. Here we report a possible association between the distribution of DSBs within higher-order chromatin structures and this pathway. Both murine severe combined immunodeficient (SCID) and Chinese hamster XR-1 cells exhibit defective DNA DSB repair and defective V(D)J recombination. The DSB repair defect is not complete, with only a subset of slowly repairing lesions affected by the mutations in these cell lines. We used a modified neutral filter elution procedure which retained elements of higher-order chromatin structures, namely nuclear matrix-DNA interactions. X-ray-induced DSBs that occurred as multiples within looped DNA structures were nonrepairable in SCID and XR-1 cells. In contrast, these lesions were repaired in radioresistant wild-type cells. Cell lines complemented with human DNA containing the respective complementing genes (XRCC7 and XRCC4) showed an increased rate of DSB repair. These results agree with previous findings with xrs5 cells (a member of the XRCC5 group). Xrs5 cells are defective for the Ku p80 subunit of the V(D)J recombination complex and show repair and V(D)J recombination defects similar to those of SCID and XR-1 cells.