Ligation of DNA after replication and repair is a prerequisite for the preservation of DNA and chromosome structure and function. Biochemical studies with Bloom's syndrome cells have revealed an abnormal DNA ligase I activity. However, genetic analysis has not revealed any differences in transcript levels or in the cDNA sequences of DNA ligase I between Bloom's syndrome and normal cells. Another human cell line, 46BR, derived from an immunodeficient patient, also has an abnormal DNA ligase I. This cell line has recently been demonstrated to harbour two different missense mutations, one at each allele of DNA ligase I. These mutations resulted in a decreased ability of partially purified cell extracts to form an enzyme-adenylate reaction intermediate. We show that 46BR hypersensitivity to an alkylating agent, ethyl methanesulphonate, and to the polyADP-ribose polymerase inhibitor 3-aminobenzamide, is rescued by transfection of wild-type DNA ligase I sequences. This provides additional genetic evidence that the defect in 46BR is at the DNA ligase I locus.