Systematic Triple-Mutant Analysis Uncovers Functional Connectivity between Pathways Involved in Chromosome Regulation

James E. Haber; Hannes Braberg; Qiuqin Wu; Richard Alexander; Julian Haase; Colm Ryan; Zach Lipkin-Moore; Kathleen E. Franks-Skiba; Tasha Johnson; Michael Shales; Tineke L. Lenstra; Frank C.P. Holstege; Jeffrey R. Johnson; Kerry Bloom; Nevan J. Krogan

doi:10.1016/j.celrep.2013.05.007

Cell Reports (Jun 2013)

Systematic Triple-Mutant Analysis Uncovers Functional Connectivity between Pathways Involved in Chromosome Regulation

James E. Haber,
Hannes Braberg,
Qiuqin Wu,
Richard Alexander,
Julian Haase,
Colm Ryan,
Zach Lipkin-Moore,
Kathleen E. Franks-Skiba,
Tasha Johnson,
Michael Shales,
Tineke L. Lenstra,
Frank C.P. Holstege,
Jeffrey R. Johnson,
Kerry Bloom,
Nevan J. Krogan

Affiliations

James E. Haber: Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Waltham, MA 02454, USA
Hannes Braberg: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Qiuqin Wu: Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Waltham, MA 02454, USA
Richard Alexander: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Julian Haase: Department of Biology, University of North Carolina, Chapel Hill, Chapel Hill, NC 27599, USA
Colm Ryan: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Zach Lipkin-Moore: Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Waltham, MA 02454, USA
Kathleen E. Franks-Skiba: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Tasha Johnson: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Michael Shales: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Tineke L. Lenstra: Molecular Cancer Research, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands
Frank C.P. Holstege: Molecular Cancer Research, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands
Jeffrey R. Johnson: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA
Kerry Bloom: Department of Biology, University of North Carolina, Chapel Hill, Chapel Hill, NC 27599, USA
Nevan J. Krogan: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94158, USA

DOI: https://doi.org/10.1016/j.celrep.2013.05.007
Journal volume & issue: Vol. 3, no. 6
pp. 2168 – 2178

Abstract

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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.

Published in Cell Reports

ISSN: 2211-1247 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: http://www.cell.com/cell-reports/home

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