Structural determinants for generating centromeric chromatin

Ben E. Black; Daniel R. Foliz; Srinivas Chakravarthy; Karolin Luger; Virgil L. Woods; Don W. Cleveland

doi:10.1038/nature02766

Structural determinants for generating centromeric chromatin

Ben E. Black, Daniel R. Foliz, Srinivas Chakravarthy, Karolin Luger, Virgil L. Woods, Don W. Cleveland^*

^*Corresponding author for this work

Biochemistry and Molecular Genetics

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

334 Scopus citations

Abstract

Mammalian centromeres are not defined by a consensus DNA sequence. In all eukaryotes a hallmark of functional centromeres-both normal ones and those formed aberrantly at atypical loci-is the accumulation of centromere protein A (CENP-A), a histone variant that replaces H3 in centromeric nucleosomes. Here we show using deuterium exchange/mass spectrometry coupled with hydrodynamic measures that CENP-A and histone H4 form sub-nucleosomal tetramers that are more compact and conformationally more rigid than the corresponding tetramers of histones H3 and H4. Substitution into histone H3 of the domain of CENP-A responsible for compaction is sufficient to direct it to centromeres. Thus, the centromere-targeting domain of CENP-A confers a unique structural rigidity to the nucleosomes into which it assembles, and is likely to have a role in maintaining centromere identity.

Original language	English (US)
Pages (from-to)	578-582
Number of pages	5
Journal	Nature
Volume	430
Issue number	6999
DOIs	https://doi.org/10.1038/nature02766
State	Published - Jul 29 2004

Funding

Acknowledgements We are indebted to E. Schwob for initiating this collaboration. We also thank A. Nakada and T. Chaplin for technical support, R. Rothstein for reagents, and J. Cau, J. Sgouros, J. Svejstrup and members of our laboratories for discussions and comments on the manuscript. A.L. was supported by an EU Marie Curie individual fellowship and a Journal of Cell Science travelling fellowship; K.S. acknowledges support through a MEXT grants-in-aid for priority areas; F.U. was supported by the EMBO Young Investigator programme. Acknowledgements We thank J. Kahana, J. Shah and K. Sullivan for reagents, and I. Cheeseman, B. Cottrell, P. Dyer, C. Gessner, F. Gordon, J. Kim, S. McBryant, D. Pantazatos and S. W. Englander for advice and technical assistance over the course of this study. We also thank J. Shah for comments on the manuscript. This research was supported by grants from the NIH to D.W.C and V.L.W., BioStar and Life Sciences Informatics grants from the University of California and ExSar Corporation to V.L.W., and by postdoctoral fellowships from the American Cancer Society (B.E.B.) and the NIH (D.R.F.). Salary support for D.W.C. was provided by the Ludwig Institute for Cancer Research.

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@article{4514bd11d940421e9c5d87b1e88345e6,

title = "Structural determinants for generating centromeric chromatin",

abstract = "Mammalian centromeres are not defined by a consensus DNA sequence. In all eukaryotes a hallmark of functional centromeres-both normal ones and those formed aberrantly at atypical loci-is the accumulation of centromere protein A (CENP-A), a histone variant that replaces H3 in centromeric nucleosomes. Here we show using deuterium exchange/mass spectrometry coupled with hydrodynamic measures that CENP-A and histone H4 form sub-nucleosomal tetramers that are more compact and conformationally more rigid than the corresponding tetramers of histones H3 and H4. Substitution into histone H3 of the domain of CENP-A responsible for compaction is sufficient to direct it to centromeres. Thus, the centromere-targeting domain of CENP-A confers a unique structural rigidity to the nucleosomes into which it assembles, and is likely to have a role in maintaining centromere identity.",

author = "Black, {Ben E.} and Foliz, {Daniel R.} and Srinivas Chakravarthy and Karolin Luger and Woods, {Virgil L.} and Cleveland, {Don W.}",

note = "Funding Information: Acknowledgements We are indebted to E. Schwob for initiating this collaboration. We also thank A. Nakada and T. Chaplin for technical support, R. Rothstein for reagents, and J. Cau, J. Sgouros, J. Svejstrup and members of our laboratories for discussions and comments on the manuscript. A.L. was supported by an EU Marie Curie individual fellowship and a Journal of Cell Science travelling fellowship; K.S. acknowledges support through a MEXT grants-in-aid for priority areas; F.U. was supported by the EMBO Young Investigator programme. Funding Information: Acknowledgements We thank J. Kahana, J. Shah and K. Sullivan for reagents, and I. Cheeseman, B. Cottrell, P. Dyer, C. Gessner, F. Gordon, J. Kim, S. McBryant, D. Pantazatos and S. W. Englander for advice and technical assistance over the course of this study. We also thank J. Shah for comments on the manuscript. This research was supported by grants from the NIH to D.W.C and V.L.W., BioStar and Life Sciences Informatics grants from the University of California and ExSar Corporation to V.L.W., and by postdoctoral fellowships from the American Cancer Society (B.E.B.) and the NIH (D.R.F.). Salary support for D.W.C. was provided by the Ludwig Institute for Cancer Research.",

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day = "29",

doi = "10.1038/nature02766",

language = "English (US)",

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T1 - Structural determinants for generating centromeric chromatin

AU - Black, Ben E.

AU - Foliz, Daniel R.

AU - Chakravarthy, Srinivas

AU - Luger, Karolin

AU - Woods, Virgil L.

AU - Cleveland, Don W.

N1 - Funding Information: Acknowledgements We are indebted to E. Schwob for initiating this collaboration. We also thank A. Nakada and T. Chaplin for technical support, R. Rothstein for reagents, and J. Cau, J. Sgouros, J. Svejstrup and members of our laboratories for discussions and comments on the manuscript. A.L. was supported by an EU Marie Curie individual fellowship and a Journal of Cell Science travelling fellowship; K.S. acknowledges support through a MEXT grants-in-aid for priority areas; F.U. was supported by the EMBO Young Investigator programme. Funding Information: Acknowledgements We thank J. Kahana, J. Shah and K. Sullivan for reagents, and I. Cheeseman, B. Cottrell, P. Dyer, C. Gessner, F. Gordon, J. Kim, S. McBryant, D. Pantazatos and S. W. Englander for advice and technical assistance over the course of this study. We also thank J. Shah for comments on the manuscript. This research was supported by grants from the NIH to D.W.C and V.L.W., BioStar and Life Sciences Informatics grants from the University of California and ExSar Corporation to V.L.W., and by postdoctoral fellowships from the American Cancer Society (B.E.B.) and the NIH (D.R.F.). Salary support for D.W.C. was provided by the Ludwig Institute for Cancer Research.

PY - 2004/7/29

Y1 - 2004/7/29

N2 - Mammalian centromeres are not defined by a consensus DNA sequence. In all eukaryotes a hallmark of functional centromeres-both normal ones and those formed aberrantly at atypical loci-is the accumulation of centromere protein A (CENP-A), a histone variant that replaces H3 in centromeric nucleosomes. Here we show using deuterium exchange/mass spectrometry coupled with hydrodynamic measures that CENP-A and histone H4 form sub-nucleosomal tetramers that are more compact and conformationally more rigid than the corresponding tetramers of histones H3 and H4. Substitution into histone H3 of the domain of CENP-A responsible for compaction is sufficient to direct it to centromeres. Thus, the centromere-targeting domain of CENP-A confers a unique structural rigidity to the nucleosomes into which it assembles, and is likely to have a role in maintaining centromere identity.

AB - Mammalian centromeres are not defined by a consensus DNA sequence. In all eukaryotes a hallmark of functional centromeres-both normal ones and those formed aberrantly at atypical loci-is the accumulation of centromere protein A (CENP-A), a histone variant that replaces H3 in centromeric nucleosomes. Here we show using deuterium exchange/mass spectrometry coupled with hydrodynamic measures that CENP-A and histone H4 form sub-nucleosomal tetramers that are more compact and conformationally more rigid than the corresponding tetramers of histones H3 and H4. Substitution into histone H3 of the domain of CENP-A responsible for compaction is sufficient to direct it to centromeres. Thus, the centromere-targeting domain of CENP-A confers a unique structural rigidity to the nucleosomes into which it assembles, and is likely to have a role in maintaining centromere identity.

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JO - Nature

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IS - 6999

ER -

Structural determinants for generating centromeric chromatin

Abstract

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