title: Systematic triple-mutant analysis uncovers functional connectivity between pathways involved in chromosome regulation.
abstract: Genetic interactions reveal the functional relationships between pairs of genes. In this study, we describe a method for the systematic generation and quantitation of triple mutants, termed triple-mutant analysis (TMA). We have used this approach to interrogate partially redundant pairs of genes in S. cerevisiae, including ASF1 and CAC1, two histone chaperones. After subjecting asf1Δ cac1Δ to TMA, we found that the Swi/Snf Rdh54 protein compensates for the absence of Asf1 and Cac1. Rdh54 more strongly associates with the chromatin apparatus and the pericentromeric region in the double mutant. Moreover, Asf1 is responsible for the synthetic lethality observed in cac1Δ strains lacking the HIRA-like proteins. A similar TMA was carried out after deleting both CLB5 and CLB6, cyclins that regulate DNA replication, revealing a strong functional connection to chromosome segregation. This approach can reveal functional redundancies that cannot be uncovered through traditional double-mutant analyses.
authors.affiliation: Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Waltham, MA 02454, USA. haber@brandeis.edu
authors.forename: James E
authors.initials: JE
authors.lastname: Haber
authors.forename: Hannes
authors.initials: H
authors.lastname: Braberg
authors.forename: Qiuqin
authors.initials: Q
authors.lastname: Wu
authors.forename: Richard
authors.initials: R
authors.lastname: Alexander
authors.forename: Julian
authors.initials: J
authors.lastname: Haase
authors.forename: Colm
authors.initials: C
authors.lastname: Ryan
authors.forename: Zach
authors.initials: Z
authors.lastname: Lipkin-Moore
authors.forename: Kathleen E
authors.initials: KE
authors.lastname: Franks-Skiba
authors.forename: Tasha
authors.initials: T
authors.lastname: Johnson
authors.forename: Michael
authors.initials: M
authors.lastname: Shales
authors.forename: Tineke L
authors.initials: TL
authors.lastname: Lenstra
authors.forename: Frank C P
authors.initials: FC
authors.lastname: Holstege
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Johnson
authors.forename: Kerry
authors.initials: K
authors.lastname: Bloom
authors.forename: Nevan J
authors.initials: NJ
authors.lastname: Krogan
mesh_terms: D002875:Chromosomes / Q000235:genetics* / Q000378:metabolism*; D004249:DNA Damage; D004261:DNA Replication; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins