Abstract

Several DNA metabolic processes are influenced or controlled by genes whose products function in DNA repair. The RAD52 gene of Saccharomyces cerevisiae plays a central role in many of the events in recombination and repair. It controls the repair of ionizing radiation-induced DNA double-strand breaks, radiation-induced spontaneous mitotic recombination, and recombination during meiosis, and is required for mutagenesis by some alkylating agents. Utilizing an antibody raised against a Neurospora crassa deoxyribonuclease, we had shown that an antigenically-related enzyme could be identified from yeast and that this enzyme is controlled by the RAD52 gene. The protein has been purified and is a single-strand exo- endonuclease and a double-strand exonuclease with MW = 72,000. It is not the product of the RAD52 gene. Using a lambda gtll vector expression library that contains genomic yeast DNA and the antibody as a probe, we have identified a segment of DNA that codes for cross-reacting material. A sequence from a yeast genomic library has been identified that hybridizes with this segment. Introduction of this into a Rad+ strain leads to enhanced synthesis of cross-reacting material. The gene appears to be essential based on mutation of the cloned sequence and introduction into the genome. The transcription of this gene is under the control of the RAD52 gene.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Intramural Research (Z01)
Project #
1Z01ES021016-08
Application #
3918607
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
8
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
U.S. National Inst of Environ Hlth Scis
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Andres, Sara N; Appel, C Denise; Westmoreland, James W et al. (2015) Tetrameric Ctp1 coordinates DNA binding and DNA bridging in DNA double-strand-break repair. Nat Struct Mol Biol 22:158-66
Ma, Wenjian; Westmoreland, Jim W; Gordenin, Dmitry A et al. (2011) Alkylation base damage is converted into repairable double-strand breaks and complex intermediates in G2 cells lacking AP endonuclease. PLoS Genet 7:e1002059
Nakai, Wataru; Westmoreland, Jim; Yeh, Elaine et al. (2011) Chromosome integrity at a double-strand break requires exonuclease 1 and MRX. DNA Repair (Amst) 10:102-10
Argueso, Juan Lucas; Westmoreland, James; Mieczkowski, Piotr A et al. (2008) Double-strand breaks associated with repetitive DNA can reshape the genome. Proc Natl Acad Sci U S A 105:11845-50
Chen, Ling; Trujillo, Kelly M; Van Komen, Stephen et al. (2005) Effect of amino acid substitutions in the rad50 ATP binding domain on DNA double strand break repair in yeast. J Biol Chem 280:2620-7
Resnick, Michael A (2005) Reduced replication: a call to ARMS. Cell 120:569-70
Lewis, L Kevin; Karthikeyan, G; Cassiano, Jared et al. (2005) Reduction of nucleosome assembly during new DNA synthesis impairs both major pathways of double-strand break repair. Nucleic Acids Res 33:4928-39
Lewis, L Kevin; Lobachev, Kirill; Westmoreland, James W et al. (2005) Use of a restriction endonuclease cytotoxicity assay to identify inducible GAL1 promoter variants with reduced basal activity. Gene 363:183-92
Lewis, L Kevin; Storici, Francesca; Van Komen, Stephen et al. (2004) Role of the nuclease activity of Saccharomyces cerevisiae Mre11 in repair of DNA double-strand breaks in mitotic cells. Genetics 166:1701-13
Lobachev, Kirill; Vitriol, Eric; Stemple, Jennifer et al. (2004) Chromosome fragmentation after induction of a double-strand break is an active process prevented by the RMX repair complex. Curr Biol 14:2107-12

Showing the most recent 10 out of 11 publications