Isolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants

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Abstract
The RAD3 gene of S. cerevisiae is required for nicking of DNA containing pyrimidine dimers or interstrand crosslinks. The RAD3 gene was cloned and physically mapped to 2.6 kilobase of DNA. A DNA segment of the cloned RAD3 insert was ligated into plasmid YIp5, which transforms yeast by homologous integration, and integrated at the RAD3 site in chromosome V, thus verifying the cloned DNA segment to be the RAD3 gene and not a suppressor. The RAD3 gene encodes a 2.5-kilobase mRNA, extending between the Kpn I site and the Sau3A1/BamHI fusion junction in plasmid pSP10, and the direction of transcription was determined. The 2.5-kilobase transcript could encode a protein of about 90,000 daltons. The deletions of the RAD3 gene are recessive lethals, indicating that the RAD3 gene plays an important role in other cellular processes in addition to incision of damaged DNA.