Defining New Functions in DNA Replication and DNA Damage Response Genes

点击翻译按钮获取中文摘要

Scientific goals: In the budding yeast S. cerevisiae, genome-wide collections of mutant strains have been employed extensively to elucidate biological function on a large scale. The resulting information has been used to place genes in pathways, to identify points of intersection between different pathways and to assign gene function to novel proteins. One flaw of these studies stems from the nature of the mutant reagents in these genome-wide collections, where gene function has been inactivated by either deletion or conditional depletion. Since many, if not all, proteins execute more than one function, such mutations are potentially pleiotropic. A potential solution is to employ separation-of-function mutations that eliminate a single biological function of a protein. However, identification of this particular class of alleles in the past has been a logistic hurdle even for single genes. This research project is based on a newly developed strategy for large-scale isolation of separation-of-function alleles which will be applied to a set of inter-related genes involved in DNA replication and response to DNA damage. Since very few mutations have been isolated for many of these genes, this research should define new functions in the DNA replication and DNA damage response pathways. Broader impacts: The scientific goals of this research project are closely intertwined with two broader impacts. First, the resulting mutations will be made widely available to the yeast community, with a particular focus on incorporation of these new reagents into genome-wide systems analysis. Genetic networks constructed from separation-of-function missense mutations are likely to uncover previously unappreciated interfaces that were missed in prior systems analysis which employed currently available mutant strains. Second, this project will rely heavily on entry-level researchers (undergraduate and high school students) who will be responsible for generating the panels of separation-of-function mutations. This experience will allow very junior researchers to play a central role in a significant research project and also introduce them to the critical function that mentoring plays in the biomedical research community.