Processing of psoralen adducts in an active human gene: repair and replication of DNA containing monoadducts and interstrand cross-links

Abstract

We have examined DNA repair in the dihydrofolate reductase (DHFR) gene in cultured human cells treated with 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) using a newly developed assay for interstrand DNA cross-linking in defined genomic sequences. Within 24 hr, 80% of the cross-links, but only 45% of the monoadducts, were removed from a 32 kb transcribed sequence, demonstrating that repair efficiency in an active human gene varies with the nature of the damage. HMT monoadducts were also detected in the replicated DHFR sequence at frequencies indicating little interference with replication. The existence of cross-linkable monoadduct sites in the replicated DNA implies strand continuity opposite those sites and a relatively error-free mechanism of bypass. Translesion replication could circumvent transcription blockage in a damaged gene. These findings have important implications for mechanisms of mutagenesis and DNA lesion tolerance in human cells.