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

被引:340
作者
Fazzio, Thomas G. [1 ]
Huff, Jason T. [1 ]
Panning, Barbara [1 ]
机构
[1] Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94158 USA
关键词
D O I
10.1016/j.cell.2008.05.031
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:162 / 174
页数:13
相关论文
共 48 条
  • [1] Integrator, a multiprotein mediator of small nuclear RNA processing, associates with the C-terminal repeat of RNA polymerase II
    Baillat, D
    Hakimi, MA
    Näär, AM
    Shilatifard, A
    Cooch, N
    Shiekhattar, R
    [J]. CELL, 2005, 123 (02) : 265 - 276
  • [2] Increased ionizing radiation sensitivity and genomic instability in the absence of histone H2AX
    Bassing, CH
    Chua, KF
    Sekiguchi, J
    Suh, H
    Whitlow, SR
    Fleming, JC
    Monroe, BC
    Ciccone, DN
    Yan, C
    Vlasakova, K
    Livingston, DM
    Ferguson, DO
    Scully, R
    Alt, FW
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (12) : 8173 - 8178
  • [3] Induction of extracellular matrix-remodeling genes by the senescence-associated protein APA-1
    Benanti, JA
    Williams, DK
    Robinson, KL
    Ozer, HL
    Galloway, DA
    [J]. MOLECULAR AND CELLULAR BIOLOGY, 2002, 22 (21) : 7385 - 7397
  • [4] A bivalent chromatin structure marks key developmental genes in embryonic stem cells
    Bernstein, BE
    Mikkelsen, TS
    Xie, XH
    Kamal, M
    Huebert, DJ
    Cuff, J
    Fry, B
    Meissner, A
    Wernig, M
    Plath, K
    Jaenisch, R
    Wagschal, A
    Feil, R
    Schreiber, SL
    Lander, ES
    [J]. CELL, 2006, 125 (02) : 315 - 326
  • [5] Polycomb complexes repress developmental regulators in murine embryonic stem cells
    Boyer, LA
    Plath, K
    Zeitlinger, J
    Brambrink, T
    Medeiros, LA
    Lee, TI
    Levine, SS
    Wernig, M
    Tajonar, A
    Ray, MK
    Bell, GW
    Otte, AP
    Vidal, M
    Gifford, DK
    Young, RA
    Jaenisch, R
    [J]. NATURE, 2006, 441 (7091) : 349 - 353
  • [6] A Brg1 null mutation in the mouse reveals functional differences among mammalian SWI/SNF complexes
    Bultman, S
    Gebuhr, T
    Yee, D
    La Mantia, C
    Nicholson, J
    Gilliam, A
    Randazzo, F
    Metzger, D
    Chambon, P
    Crabtree, G
    Magnuson, T
    [J]. MOLECULAR CELL, 2000, 6 (06) : 1287 - 1295
  • [7] Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells
    Chambers, I
    Colby, D
    Robertson, M
    Nichols, J
    Lee, S
    Tweedie, S
    Smith, A
    [J]. CELL, 2003, 113 (05) : 643 - 655
  • [8] Nanog safeguards pluripotency and mediates germline development
    Chambers, Ian
    Silva, Jose
    Colby, Douglas
    Nichols, Jennifer
    Nijmeijer, Bianca
    Robertson, Morag
    Vrana, Jan
    Jones, Ken
    Grotewold, Lars
    Smith, Austin
    [J]. NATURE, 2007, 450 (7173) : 1230 - U8
  • [9] The p400 E1A-associated protein is a novel component of the p53→p21 senescence pathway
    Chan, HM
    Narita, M
    Lowe, SW
    Livingstone, DM
    [J]. GENES & DEVELOPMENT, 2005, 19 (02) : 196 - 201
  • [10] Minichromosome maintenance proteins are direct targets of the ATM and ATR checkpoint kinases
    Cortez, D
    Glick, G
    Elledge, SJ
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (27) : 10078 - 10083