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Characterization of genes that are synthetically lethal with ade3 or leu2 in Saccharomyces cerevisiae
Author(s) -
Nigavekar Shraddha S.,
Can John F.
Publication year - 2001
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.807
Subject(s) - mutant , biology , plasmid , gene , lethal allele , mutation , genetic screen , genetics , phenotype , synthetic lethality
Combinations of two non‐lethal mutations that result in cell death are synthetically lethal. Such a genetic relationship suggests a functional interaction between the corresponding gene products. Frequently, an ade2 ade3 colony‐sectoring assay is used to screen for synthetic lethal mutants. In these screens, mutants are sought that fail to lose a plasmid that bears a gene of interest. However, a subset of mutants is often found that is dependent on plasmid components other than the target gene. To understand the mechanism of this dependence, we characterized those mutants that, although prevalent in most mutant hunts, are usually discarded. Using a LEU2–ADE3 plasmid, plasmid‐dependent mutations were found in the SHM2 , PTR3 , BAP2 and SSY1 genes. Double shm2 ade3 mutants are non‐viable because the two pathways for tetrahydrofolate synthesis are blocked. Mutations in PTR3 , BAP2 and SSY1 disrupt sensing and transport of extracellular leucine. Therefore, ptr3 , bap2 or ssy1 mutants must be leucine prototrophs to grow on rich media. In light of these findings, we propose modifications that should improve the efficiency of synthetic lethal screening procedures. Copyright © 2002 John Wiley & Sons, Ltd.