Characterization of HRD3, a Schizosaccharomyces pombe gene involved in DNA repair and cell viability

The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. The RAD3 encoded protein possesses a single stranded DNA‐dependent ATPase and DNA and DNA‐RNA helicase activities. To examine the extent of conservation of structure and function of a S. pombe RAD3 during eukaryotic evolution, the RAD3 homolog gene was isolated by screening of genomic DNA library. The isolated gene was designated as HRD3 (homolog of RAD3 gene). Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as HRD3 gene and this gene exists as a single copy in S. pombe. The transcript of 2. 8 kb was detected by Northern blot analysis. The level of transcripts increased by ultraviolet (UV) irradiation, indicating that HRD3 is one of the UV‐inducible genes in S. pombe. Furthermore, the predicted partial sequence of HRD3 protein has 60% identity to S. cerevisiae RAD3 gene. This homology was particularly striking in the regions identified as being conserved in a group of DNA helicases. Gene deletion experiments indicate that the HRD3 gene is essential for viability and DNA repair function. These observations suggest evolutionary conservation of other protein components with which HRD3 might interact in mediating its DNA repair and viability functions.