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PMID: 18614019 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

An RNAi screen of chromatin proteins identifies Tip60-p400 as a regulator of embryonic stem cell identity.

Cell ·Vol. 134 ·No. 1 ·2008-07-11 ·Pages 162-74

Fazzio TG, Huff JT, Panning B

Abstract

Proper regulation of chromatin structure is necessary for the maintenance of cell type-specific gene expression patterns. The embryonic stem cell (ESC) expression pattern governs self-renewal and pluripotency. Here, we present an RNAi screen in mouse ESCs of 1008 loci encoding chromatin proteins. We identified 68 proteins that exhibit diverse phenotypes upon knockdown (KD), including seven subunits of the Tip60-p400 complex. Phenotypic analyses revealed that Tip60-p400 is necessary to maintain characteristic features of ESCs. We show that p400 localization to the promoters of both silent and active genes is dependent upon histone H3 lysine 4 trimethylation (H3K4me3). Furthermore, the Tip60-p400 KD gene expression profile is enriched for developmental regulators and significantly overlaps with that of the transcription factor Nanog. Depletion of Nanog reduces p400 binding to target promoters without affecting H3K4me3 levels. Together, these data indicate that Tip60-p400 integrates signals from Nanog and H3K4me3 to regulate gene expression in ESCs.

MeSH Terms
Animals Calcium Channels/metabolism Embryonic Stem Cells/cytology,metabolism Histone Acetyltransferases/metabolism Histones/metabolism Homeodomain Proteins/metabolism Inositol 1,4,5-Trisphosphate Receptors Lysine Acetyltransferase 5 Mice Nanog Homeobox Protein Promoter Regions, Genetic RNA Interference Receptors, Cytoplasmic and Nuclear/metabolism Trans-Activators Transcription, Genetic
Chemicals
Calcium Channels Histones Homeodomain Proteins Inositol 1,4,5-Trisphosphate Receptors Itpr1 protein, mouse Nanog Homeobox Protein Nanog protein, mouse Receptors, Cytoplasmic and Nuclear Trans-Activators Histone Acetyltransferases Kat5 protein, mouse Lysine Acetyltransferase 5
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fazzio Thomas G
Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94158, USA. [email protected]
Huff Jason T
Panning Barbara
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Article Info
Journal
Cell
Abbr.
Cell
ISSN
1097-4172
DOI
10.1016/j.cell.2008.05.031
Email
[email protected]
Published
2008-07-11
Pages
162-74
Language
English
Region
United States
NLM ID
0413066
PMCID
PMC4308735
Subset
IM
Grants
NIGMS NIH HHS · R01 GM063671 · United States
NIGMS NIH HHS · R01 GM085186 · United States
NIGMS NIH HHS · R01 GM085186-01A1 · United States
NIGMS NIH HHS · R01 GM63671 · United States
Databases
GEO
GSE11243
External Links
PubMed source DOI: 10.1016/j.cell.2008.05.031 PMC Full Text View journal details
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