Identification and expression of the gene product encoding a CPD photolyase from Dunaliella salina

Abstract: Ultraviolet light induces photoproducts, cyclobutane pyrimidine dimers (CPDs) and (6-4) photoproducts (6-4PPs), in cellular DNA, which cause cytotoxic and genotoxic effects on the cells. Cells have several DNA repair mechanisms to repair the damage and to maintain genetic information of the cells. Photoreactivation is one of the DNA repair mechanism to remove UV-induced DNA damage from cellular DNA catalyzed by photolyase under visible light. Two types of photolyase, CPD photolyase and (6-4) photolyase, are specific for CPDs and for (6-4)PPs. We have isolated a gene product encoding CPD photolyase, named PHR2, from Dunaliella salina which is a kind of unicellular alga. Sequence analysis showed that PHR2 encodes a protein that has 529 amino acids and is similar to other Class II CPD photolyase. The complementation assay of the photoreactivation deficiency of the Escherichia coli SY2 by PHR2 cDNA showed a significant increase in survival rate when cells were irradiated with UV-C. Real-time PCR analysis indicated that the transcription of PHR2 was induced by UV-C, white light, high salinity, and H2O2. [Copyright &y& Elsevier]