The recognition and removal of interstrand cross-links is perhaps the least understood of all repair pathways in eukaryotic cells. We have shown previously that uncoupling of cross-links occurs in mammalian cell extracts and have identified a number of factors that mediate this process. However, we have not observed complete repair of the substrate in this system. Here, we show that uncoupling of interstrand cross-links also occurs in Xenopus laevis egg extracts, and that the initial products of this reaction are identical to the products observed in mammalian cell extracts suggesting a common mechanism. However in contrast to mammalian cell extracts, we observe repair of the cross-linked substrate in the Xenopus extracts presumably by a translesion bypass mechanism that allows replication past the uncoupled monoadduct, and its likely,subsequent removal by nucleotide excision repair. This repair process is shown to be highly mutagenic consistent with bypass synthesis. (c) 2005 Elsevier Inc. All rights reserved.