DNA and Chromatin Modification Networks Distinguish Stem Cell Pluripotent Ground States

October 1, 2012

Pluripotent stem cells are capable of differentiating into all cell types of the body and therefore hold tremendous promise for regenerative medicine. Despite their widespread use in laboratories across the world, a detailed understanding of the molecular mechanisms that regulate the pluripotent state is currently lacking. Mouse embryonic (mESC) and epiblast (mEpiSC) stem cells are two closely related classes of pluripotent stem cells, derived from distinct embryonic tissues. Although both mESC and mEpiSC are pluripotent, these cell types show important differences in their properties suggesting distinct pluripotent ground states. To understand the molecular basis of pluripotency, we analyzed the nuclear proteomes of mESCs and mEpiSCs to identify protein networks that regulate their respective pluripotent states. Our study used label-free LC-MS/MS to identify and quantify 1597 proteins in embryonic and epiblast stem cell nuclei. Immunoblotting of a selected protein subset was used to confirm that key components of chromatin regulatory networks are differentially expressed in mESCs and mEpiSCs. Specifically, we identify differential expression of DNA methylation, ATP-dependent chromatin remodeling and nucleosome remodeling networks in mESC and mEpiSC nuclei. This study is the first comparative study of protein networks in cells representing the two distinct, pluripotent states, and points to the importance of DNA and chromatin modification processes in regulating pluripotency. In addition, by integrating our data with existing pluripotency networks, we provide detailed maps of protein networks that regulate pluripotency that will further both the fundamental understanding of pluripotency as well as efforts to reliably control the differentiation of these cells into functional cell fates.

Figure Selected mESC- and mEpiSC-associated protein networks. Ingenuity Pathway Analysis (IPA) reveals differentially expressed subnetworks including (A) Polycomb-group (PcG), DNA methyltransferase, Nucleosome remodeling/histone deacetylase (NuRD), OCT4-SOX2-NANOG and SWI/SNF BAF complexes, B, Protein networks functioning in DNA replication and Genome surveillance/DNA repair, (C) Transcription factor complexes, and (D) Homeobox protein complex. All proteins shown were identified as being either more abundant in mESC (red nodes) or more abundant in mEpiSC (green nodes) at p < 0.1 (Student's t test).

Results from: Song, J.,Saha, S., Gokulrangan, G., Tesar, P. J. and Ewing, R.M.. Molecular & Cellular Proteomics, 11, 1036-1047. October 1, 2012.