Genetic and biochemical analysis of photolyase mutants ofChlamydomonas reinhardtii

Abstract

A mutant ofChlamydomonas reinhardtii phr-1-1 is deficient in the photorepair of pyrimidine dimers in nuclear DNA but not in chloroplast DNA. In this report, a second photoreactivation-deficient strain, phr-1-2, which has a similar phenotype as phr-1-1, is described. To determine if these mutations were in different genes, complementation tests as well as tetrad analysis were performed. Six diploid strains were constructed. The diploid strains containing one mutation exhibited similar UV-light survival curves under photoreactivating conditions as the wild-type diploid, indicating recessive nature of the mutations. No increase in survival was obtained with phr-1-1 phr-1-2 compared with either the phr-1-1 phr-1-1 or phr-1-2 phr-1-2 diploids indicating a lack of complementation. The amount of DNA photolyase activity in cell-free extracts of diploids with one mutation was not significantly different from extracts of wild-type diploids indicating a lack of gene dosage. The amount of DNA photolyase activity in extracts from the phr-1-1 phr-1-2 was no greater than found in the phr-1-1 phr-1-1 or phr-1-2 phr-1-2 diploids, confirming a lack of complementation of the mutations. Analysis of 106 tetrads from a cross ofphr-1-1 arg2×phr-1-2 arg7 indicated thatphr-1-1 andphr-1-2 were mutations in the same gene.