Open AccessIsolation and characterization of the RAD3 gene of Saccharomyces cerevisiae and inviability of rad3 deletion mutants
Author(s) -
David R. Higgins,
Satya Prakash,
Paul Reynolds,
Renata Polakowska,
Shane Weber,
Louise Prakash
Publication year - 1983
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.80.18.5680
Subject(s) - biology , gene , microbiology and biotechnology , plasmid , dna , saccharomyces cerevisiae , genetics
TheRAD3 gene ofSaccharomyces cerevisiae is required for nicking of DNA containing pyrimidine dimers or interstrand crosslinks. We have cloned theRAD3 gene and physically mapped it to 2.6 kilobase of DNA. A DNA segment of the clonedRAD3 insert was ligated into plasmid YIp5, which transforms yeast by homologous integration, and shown to integrate at theRAD3 site in chromosome V, thus verifying the cloned DNA segment to be theRAD3 gene and not a suppressor. TheRAD3 gene encodes a 2.5-kilobase mRNA, extending between theKpn I site and theSau 3A1/Bam HI fusion junction in plasmid pSP10, and the direction of transcription has been determined. The 2.5-kilobase transcript could encode a protein of about 90,000 daltons. We also show the deletions of theRAD3 gene to be recessive lethals, indicating that theRAD3 gene plays an important role in other cellular processes in addition to incision of damaged DNA.