Functional and mechanistic studies of XPC DNA-repair complex as transcriptional coactivator in embryonic stem cells

Proc Natl Acad Sci U S A. 2015 May 5;112(18):E2317-26. doi: 10.1073/pnas.1505569112. Epub 2015 Apr 21.

Abstract

The embryonic stem cell (ESC) state is transcriptionally controlled by OCT4, SOX2, and NANOG with cofactors, chromatin regulators, noncoding RNAs, and other effectors of signaling pathways. Uncovering components of these regulatory circuits and their interplay provides the knowledge base to deploy ESCs and induced pluripotent stem cells. We recently identified the DNA-repair complex xeroderma pigmentosum C (XPC)-RAD23B-CETN2 as a stem cell coactivator (SCC) required for OCT4/SOX2 transcriptional activation. Here we investigate the role of SCC genome-wide in murine ESCs by mapping regions bound by RAD23B and analyzing transcriptional profiles of SCC-depleted ESCs. We establish OCT4 and SOX2 as the primary transcription factors recruiting SCC to regulatory regions of pluripotency genes and identify the XPC subunit as essential for interaction with the two proteins. The present study reveals new mechanistic and functional aspects of SCC transcriptional activity, and thus underscores the diversified functions of this regulatory complex.

Keywords: ChIP-seq; RNA-seq; pluripotency; protein–protein interactions; transcription.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Binding Sites
  • Cell Differentiation
  • Cell Lineage
  • DNA Repair
  • DNA-Binding Proteins / metabolism*
  • Embryonic Stem Cells / cytology*
  • Gene Expression Regulation, Developmental*
  • Genome
  • HEK293 Cells
  • Humans
  • Immunoglobulin G / chemistry
  • Lentivirus / metabolism
  • Mice
  • Mice, Knockout
  • Pluripotent Stem Cells / cytology
  • Promoter Regions, Genetic
  • Protein Binding
  • SOXB1 Transcription Factors / metabolism
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • DNA-Binding Proteins
  • Immunoglobulin G
  • Rad23b protein, mouse
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Transcription Factors
  • XPC protein, human

Associated data

  • GEO/GSE64040