UMMS Affiliation

Program in Gene Function and Expression; Program in Molecular Medicine

Date

11-13-2012

Document Type

Article

Medical Subject Headings

TATA-Box Binding Protein; TATA-Binding Protein Associated Factors; Transcription Factors; Transcription Factors, General; Embryonic Stem Cells

Disciplines

Genetics and Genomics

Abstract

The general transcription factor TFIID comprises the TATA-box-binding protein (TBP) and approximately 14 TBP-associated factors (TAFs). Here we find, unexpectedly, that undifferentiated human embryonic stem cells (hESCs) contain only six TAFs (TAFs 2, 3, 5, 6, 7 and 11), whereas following differentiation all TAFs are expressed. Directed and global chromatin immunoprecipitation analyses reveal an unprecedented promoter occupancy pattern: most active genes are bound by only TAFs 3 and 5 along with TBP, whereas the remaining active genes are bound by TBP and all six hESC TAFs. Consistent with these results, hESCs contain a previously undescribed complex comprising TAFs 2, 6, 7, 11 and TBP. Altering the composition of hESC TAFs, either by depleting TAFs that are present or ectopically expressing TAFs that are absent, results in misregulated expression of pluripotency genes and induction of differentiation. Thus, the selective expression and use of TAFs underlies the ability of hESCs to self-renew.

Comments

Citation: Maston et al. eLife 2012;1:e00068. DOI: 10.7554/eLife.00068. Link to article on publisher's site

Copyright Maston et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

Supplementary file is also available at http://dx.doi.org/10.7554/eLife.00068.026.

The following datasets were generated: Maston GA, Green MR, Zhu LJ, Chamberlain L, Lin L, 2012, ChIP-chip from H9 hESCs for RNA Polymerase II, TBP, TAF3, TAF5, TAF7, and TAF11, GSE39312; http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE39312, In the public domain at GEO: http://www.ncbi.nlm.nih.gov/geo/.

Related Resources

Link to article in PubMed

Keywords

datasets

PubMed ID

23150797

elife00068s001.xls (35 kB)
Supplementary file 1. List of antibodies, primer sequences and siRNA/shRNAs used in this study. (A) List of antibodies used in this study. (B) Primers sequences for qRT-PCR, ChIP and ChIP-chip validation experiments. (C) Sequences of synthesized siRNAs, and clone ID numbers for shRNAs obtained from Open Biosystems (the luciferase siRNA sequence was previously reported (Elbashir et al., 2001)).

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