A Functional Genomics Approach to Identify Novel Breast Cancer Gene Targets in Yeast
Abstract
We are using the yeast Saccharomyces cerevisiae to identify new cancer gene targets that interact with the tumor suppressor Brcal. Expression of Ercal in diploid WT yeast leads to prolonged Gi arrest and lethality. We identified from a collection of ionizing radiation (IR) -sensitive yeast deletion strains or from a pool of 4%46 genetically tagged deletion strains, 34 that rescue Brcal-induced lethality. Two IR resistance genes that rescue Brcal-induced lethality are the transcription factors CCR4 and DHHl. These are checkpoint genes required for cell cycle progression in Gl and S phases following DNA damage. Consistent with a role in radiation resistance, Dhhlp and its highly conserved human ortholog DDX6 were found to physically interact with Brcal in yeast and human cells. Another transcription factor (YAF9) was IR sensitive and rescued Brcal-induced lethality when deleted. This deletion strain and 19 others were subsequently isolated from the deletion strain pool. Most of these deletions (75%) were IR sensitive and hypersensitive to the toxin zymocin which appears to induce DSB damage by inhibiting transcription. Furthermore, most (85%) of these genes are highly conserved suggesting that the hunan orthologs may interact with Brcal to maintain genomic stability and suppress the onset of breast cancer.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2004
- Accession Number
- ADA432231
Entities
People
- Craig Bennett
Organizations
- Duke University