Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants

Junbiao Dai; Edel M. Hyland; Daniel S. Yuan; Hailiang Huang; Joel S. Bader; Jef D. Boeke

doi:10.1016/j.cell.2008.07.019

Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants

Junbiao Dai, Edel M. Hyland, Daniel S. Yuan, Hailiang Huang, Joel S. Bader, Jef D. Boeke

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

Abstract

Nucleosome structural integrity underlies the regulation of DNA metabolism and transcription. Using a synthetic approach, a versatile library of 486 systematic histone H3 and H4 substitution and deletion mutants that probes the contribution of each residue to nucleosome function was generated in Saccharomyces cerevisiae. We probed fitness contributions of each residue to perturbations of chromosome integrity and transcription, mapping global patterns of chemical sensitivities and requirements for transcriptional silencing onto the nucleosome surface. Each histone mutant was tagged with unique molecular barcodes, facilitating identification of histone mutant pools through barcode amplification, labeling, and TAG microarray hybridization. Barcodes were used to score complex phenotypes such as competitive fitness in a chemostat, DNA repair proficiency, and synthetic genetic interactions, revealing new functions for distinct histone residues and new interdependencies among nucleosome components and their modifiers.

Original language	English (US)
Pages (from-to)	1066-1078
Number of pages	13
Journal	Cell
Volume	134
Issue number	6
DOIs	https://doi.org/10.1016/j.cell.2008.07.019
State	Published - Sep 19 2008

Keywords

CHEMBIO
PROTEINS

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.cell.2008.07.019

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title = "Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants",

abstract = "Nucleosome structural integrity underlies the regulation of DNA metabolism and transcription. Using a synthetic approach, a versatile library of 486 systematic histone H3 and H4 substitution and deletion mutants that probes the contribution of each residue to nucleosome function was generated in Saccharomyces cerevisiae. We probed fitness contributions of each residue to perturbations of chromosome integrity and transcription, mapping global patterns of chemical sensitivities and requirements for transcriptional silencing onto the nucleosome surface. Each histone mutant was tagged with unique molecular barcodes, facilitating identification of histone mutant pools through barcode amplification, labeling, and TAG microarray hybridization. Barcodes were used to score complex phenotypes such as competitive fitness in a chemostat, DNA repair proficiency, and synthetic genetic interactions, revealing new functions for distinct histone residues and new interdependencies among nucleosome components and their modifiers.",

keywords = "CHEMBIO, PROTEINS",

author = "Junbiao Dai and Hyland, {Edel M.} and Yuan, {Daniel S.} and Hailiang Huang and Bader, {Joel S.} and Boeke, {Jef D.}",

note = "Funding Information: J.D. and E.M.H. contributed equally and are listed alphabetically. We thank Jeffry Corden and Lisa Scheifele for help with chemostats, Anne Norris for discussions and assistance with molecular graphics, and Lisa Chengran Huang, Qing Huang, and Yu-yi Lin for assistance with phenotypic screening. We thank Alain Verreault and Yi Zhang for providing antibodies. This work was supported in part by NIH Roadmap grant U54 RR020839 to J.S.B. and J.D.B. ",

year = "2008",

month = sep,

day = "19",

doi = "10.1016/j.cell.2008.07.019",

language = "English (US)",

volume = "134",

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T1 - Probing Nucleosome Function

T2 - A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants

AU - Dai, Junbiao

AU - Hyland, Edel M.

AU - Yuan, Daniel S.

AU - Huang, Hailiang

AU - Bader, Joel S.

AU - Boeke, Jef D.

N1 - Funding Information: J.D. and E.M.H. contributed equally and are listed alphabetically. We thank Jeffry Corden and Lisa Scheifele for help with chemostats, Anne Norris for discussions and assistance with molecular graphics, and Lisa Chengran Huang, Qing Huang, and Yu-yi Lin for assistance with phenotypic screening. We thank Alain Verreault and Yi Zhang for providing antibodies. This work was supported in part by NIH Roadmap grant U54 RR020839 to J.S.B. and J.D.B.

PY - 2008/9/19

Y1 - 2008/9/19

N2 - Nucleosome structural integrity underlies the regulation of DNA metabolism and transcription. Using a synthetic approach, a versatile library of 486 systematic histone H3 and H4 substitution and deletion mutants that probes the contribution of each residue to nucleosome function was generated in Saccharomyces cerevisiae. We probed fitness contributions of each residue to perturbations of chromosome integrity and transcription, mapping global patterns of chemical sensitivities and requirements for transcriptional silencing onto the nucleosome surface. Each histone mutant was tagged with unique molecular barcodes, facilitating identification of histone mutant pools through barcode amplification, labeling, and TAG microarray hybridization. Barcodes were used to score complex phenotypes such as competitive fitness in a chemostat, DNA repair proficiency, and synthetic genetic interactions, revealing new functions for distinct histone residues and new interdependencies among nucleosome components and their modifiers.

AB - Nucleosome structural integrity underlies the regulation of DNA metabolism and transcription. Using a synthetic approach, a versatile library of 486 systematic histone H3 and H4 substitution and deletion mutants that probes the contribution of each residue to nucleosome function was generated in Saccharomyces cerevisiae. We probed fitness contributions of each residue to perturbations of chromosome integrity and transcription, mapping global patterns of chemical sensitivities and requirements for transcriptional silencing onto the nucleosome surface. Each histone mutant was tagged with unique molecular barcodes, facilitating identification of histone mutant pools through barcode amplification, labeling, and TAG microarray hybridization. Barcodes were used to score complex phenotypes such as competitive fitness in a chemostat, DNA repair proficiency, and synthetic genetic interactions, revealing new functions for distinct histone residues and new interdependencies among nucleosome components and their modifiers.

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KW - PROTEINS

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Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants

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Keywords

ASJC Scopus subject areas

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