Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal

Gundula Streubel; Darren J. Fitzpatrick; Giorgio Oliviero; Andrea Scelfo; Bruce Moran; Sudipto Das; Nayla Munawar; Ariane Watson; Kieran Wynne; Gian Luca Negri; Eugene T. Dillon; Sri Ganesh Jammula; Karsten Hokamp; Darran P. O'Connor; Diego Pasini; Gerard Cagney; Adrian P. Bracken

doi:10.15252/embj.201696307

Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal

Gundula Streubel, Darren J. Fitzpatrick, Giorgio Oliviero, Andrea Scelfo, Bruce Moran, Sudipto Das, Nayla Munawar, Ariane Watson, Kieran Wynne, Gian Luca Negri, Eugene T. Dillon, Sri Ganesh Jammula, Karsten Hokamp, Darran P. O'Connor, Diego Pasini, Gerard Cagney, Adrian P. Bracken

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

28 Citations (Scopus)

Abstract

Sin3a is the central scaffold protein of the prototypical Hdac1/2 chromatin repressor complex, crucially required during early embryonic development for the growth of pluripotent cells of the inner cell mass. Here, we compare the composition of the Sin3a-Hdac complex between pluripotent embryonic stem (ES) and differentiated cells by establishing a method that couples two independent endogenous immunoprecipitations with quantitative mass spectrometry. We define the precise composition of the Sin3a complex in multiple cell types and identify the Fam60a subunit as a key defining feature of a variant Sin3a complex present in ES cells, which also contains Ogt and Tet1. Fam60a binds on H3K4me3-positive promoters in ES cells, together with Ogt, Tet1 and Sin3a, and is essential to maintain the complex on chromatin. Finally, we show that depletion of Fam60a phenocopies the loss of Sin3a, leading to reduced proliferation, an extended G1-phase and the deregulation of lineage genes. Taken together, Fam60a is an essential core subunit of a variant Sin3a complex in ES cells that is required to promote rapid proliferation and prevent unscheduled differentiation.

Original language	English
Pages (from-to)	2216-2232
Number of pages	17
Journal	EMBO Journal
Volume	36
Issue number	15
DOIs	https://doi.org/10.15252/embj.201696307
Publication status	Published - Aug 1 2017
Externally published	Yes

Keywords

Fam60a
Sin3a-Hdac complex
embryonic stem cell
self-renewal

ASJC Scopus subject areas

Neuroscience(all)
Molecular Biology
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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10.15252/embj.201696307

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Streubel, G., Fitzpatrick, D. J., Oliviero, G., Scelfo, A., Moran, B., Das, S., Munawar, N., Watson, A., Wynne, K., Negri, G. L., Dillon, E. T., Jammula, S. G., Hokamp, K., O'Connor, D. P., Pasini, D., Cagney, G., & Bracken, A. P. (2017). Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal. EMBO Journal, 36(15), 2216-2232. https://doi.org/10.15252/embj.201696307

Streubel, G, Fitzpatrick, DJ, Oliviero, G, Scelfo, A, Moran, B, Das, S, Munawar, N, Watson, A, Wynne, K, Negri, GL, Dillon, ET, Jammula, SG, Hokamp, K, O'Connor, DP, Pasini, D, Cagney, G & Bracken, AP 2017, 'Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal', EMBO Journal, vol. 36, no. 15, pp. 2216-2232. https://doi.org/10.15252/embj.201696307

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title = "Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal",

abstract = "Sin3a is the central scaffold protein of the prototypical Hdac1/2 chromatin repressor complex, crucially required during early embryonic development for the growth of pluripotent cells of the inner cell mass. Here, we compare the composition of the Sin3a-Hdac complex between pluripotent embryonic stem (ES) and differentiated cells by establishing a method that couples two independent endogenous immunoprecipitations with quantitative mass spectrometry. We define the precise composition of the Sin3a complex in multiple cell types and identify the Fam60a subunit as a key defining feature of a variant Sin3a complex present in ES cells, which also contains Ogt and Tet1. Fam60a binds on H3K4me3-positive promoters in ES cells, together with Ogt, Tet1 and Sin3a, and is essential to maintain the complex on chromatin. Finally, we show that depletion of Fam60a phenocopies the loss of Sin3a, leading to reduced proliferation, an extended G1-phase and the deregulation of lineage genes. Taken together, Fam60a is an essential core subunit of a variant Sin3a complex in ES cells that is required to promote rapid proliferation and prevent unscheduled differentiation.",

keywords = "Fam60a, Sin3a-Hdac complex, embryonic stem cell, self-renewal",

author = "Gundula Streubel and Fitzpatrick, {Darren J.} and Giorgio Oliviero and Andrea Scelfo and Bruce Moran and Sudipto Das and Nayla Munawar and Ariane Watson and Kieran Wynne and Negri, {Gian Luca} and Dillon, {Eugene T.} and Jammula, {Sri Ganesh} and Karsten Hokamp and O'Connor, {Darran P.} and Diego Pasini and Gerard Cagney and Bracken, {Adrian P.}",

note = "Funding Information: We thank members of the Bracken laboratory for helpful discussions and critical reading of the manuscript. We also thank Dr. John Rouse, University of Dundee, for providing the Fam60a antibody. We thank members of the Pasini Lab for the helpful support and Rut Klinger for help with the RNA-Seq sample preparation. S.D was supported by the Irish Cancer Society post-doctoral fellowship—CRF13DAS. Work in the Cagney Lab was supported by Science Foundation Ireland (SFI/10/1N.1/B3.19). Work in the Bracken Lab is supported by Science Foundation Ireland under the Principal Investigator Career Advancement Award (SFI/10/IN.1/B3002), the Health Research Board under the Health Research Awards 2010 (HRA_POR/2010/124), the Irish Research Council, St. Vincent's Foundation and the Irish Cancer Society Collaborative Cancer Research Centre, and BREAST-PREDICT Grant, CCRC13GAL (www.breastpredict.com). Publisher Copyright: {\textcopyright} 2017 The Authors",

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doi = "10.15252/embj.201696307",

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T1 - Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal

AU - Streubel, Gundula

AU - Fitzpatrick, Darren J.

AU - Oliviero, Giorgio

AU - Scelfo, Andrea

AU - Moran, Bruce

AU - Das, Sudipto

AU - Munawar, Nayla

AU - Watson, Ariane

AU - Wynne, Kieran

AU - Negri, Gian Luca

AU - Dillon, Eugene T.

AU - Jammula, Sri Ganesh

AU - Hokamp, Karsten

AU - O'Connor, Darran P.

AU - Pasini, Diego

AU - Cagney, Gerard

AU - Bracken, Adrian P.

N1 - Funding Information: We thank members of the Bracken laboratory for helpful discussions and critical reading of the manuscript. We also thank Dr. John Rouse, University of Dundee, for providing the Fam60a antibody. We thank members of the Pasini Lab for the helpful support and Rut Klinger for help with the RNA-Seq sample preparation. S.D was supported by the Irish Cancer Society post-doctoral fellowship—CRF13DAS. Work in the Cagney Lab was supported by Science Foundation Ireland (SFI/10/1N.1/B3.19). Work in the Bracken Lab is supported by Science Foundation Ireland under the Principal Investigator Career Advancement Award (SFI/10/IN.1/B3002), the Health Research Board under the Health Research Awards 2010 (HRA_POR/2010/124), the Irish Research Council, St. Vincent's Foundation and the Irish Cancer Society Collaborative Cancer Research Centre, and BREAST-PREDICT Grant, CCRC13GAL (www.breastpredict.com). Publisher Copyright: © 2017 The Authors

PY - 2017/8/1

Y1 - 2017/8/1

N2 - Sin3a is the central scaffold protein of the prototypical Hdac1/2 chromatin repressor complex, crucially required during early embryonic development for the growth of pluripotent cells of the inner cell mass. Here, we compare the composition of the Sin3a-Hdac complex between pluripotent embryonic stem (ES) and differentiated cells by establishing a method that couples two independent endogenous immunoprecipitations with quantitative mass spectrometry. We define the precise composition of the Sin3a complex in multiple cell types and identify the Fam60a subunit as a key defining feature of a variant Sin3a complex present in ES cells, which also contains Ogt and Tet1. Fam60a binds on H3K4me3-positive promoters in ES cells, together with Ogt, Tet1 and Sin3a, and is essential to maintain the complex on chromatin. Finally, we show that depletion of Fam60a phenocopies the loss of Sin3a, leading to reduced proliferation, an extended G1-phase and the deregulation of lineage genes. Taken together, Fam60a is an essential core subunit of a variant Sin3a complex in ES cells that is required to promote rapid proliferation and prevent unscheduled differentiation.

AB - Sin3a is the central scaffold protein of the prototypical Hdac1/2 chromatin repressor complex, crucially required during early embryonic development for the growth of pluripotent cells of the inner cell mass. Here, we compare the composition of the Sin3a-Hdac complex between pluripotent embryonic stem (ES) and differentiated cells by establishing a method that couples two independent endogenous immunoprecipitations with quantitative mass spectrometry. We define the precise composition of the Sin3a complex in multiple cell types and identify the Fam60a subunit as a key defining feature of a variant Sin3a complex present in ES cells, which also contains Ogt and Tet1. Fam60a binds on H3K4me3-positive promoters in ES cells, together with Ogt, Tet1 and Sin3a, and is essential to maintain the complex on chromatin. Finally, we show that depletion of Fam60a phenocopies the loss of Sin3a, leading to reduced proliferation, an extended G1-phase and the deregulation of lineage genes. Taken together, Fam60a is an essential core subunit of a variant Sin3a complex in ES cells that is required to promote rapid proliferation and prevent unscheduled differentiation.

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Fam60a defines a variant Sin3a-Hdac complex in embryonic stem cells required for self-renewal

Abstract

Keywords

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