UMMS Affiliation
Program in Gene Function and Expression; Program in Molecular Medicine; Program in Bioinformatics and Integrative Biology
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.
Related Resources
Keywords
datasets
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)).
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/.