Single-molecule analysis reveals changes in the DNA replication program for the POU5F1 locus upon human embryonic stem cell differentiation

Sherri S. Schultz, Sabrina C. Desbordes, Zhuo Du, Settapong Kosiyatrakul, Inna Lipchina, Lorenz Studer, Carl L. Schildkraut

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Human embryonic stem cells (hESCs), due to their pluripotent nature, represent a particularly relevant model system to study the relationship between the replication program and differentiation state. Here, we define the basic properties of the replication program in hESCs and compare them to the programs of hESC-derived multipotent cells (neural rosette cells) and primary differentiated cells (microvascular endothelial cells [MECs]). We characterized three genomic loci: two pluripotency regulatory genes, POU5F1 (OCT4) and NANOG, and the IGH locus, a locus that is transcriptionally active specifically in B-lineage cells. We applied a high-resolution approach to capture images of individual replicated DNA molecules. We demonstrate that for the loci studied, several basic properties of replication, including the average speed of replication forks and the average density of initiation sites, were conserved among the cells analyzed. We also demonstrate, for the first time, the presence of initiation zones in hESCs. However, significant differences were evident in other aspects of replication for the DNA segment containing the POU5F1 gene. Specifically, the locations of centers of initiation zones and the direction of replication fork progression through the POU5F1 gene were conserved in two independent hESC lines but were different in hESC-derived multipotent cells and MECs. Thus, our data identify features of the replication program characteristic of hESCs and define specific changes in replication during hESC differentiation.

Original languageEnglish (US)
Pages (from-to)4521-4534
Number of pages14
JournalMolecular and Cellular Biology
Volume30
Issue number18
DOIs
StatePublished - 2010

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DNA Replication
Cell Differentiation
Endothelial Cells
Human Embryonic Stem Cells
Regulator Genes
Genes
Cell Line
DNA

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Single-molecule analysis reveals changes in the DNA replication program for the POU5F1 locus upon human embryonic stem cell differentiation. / Schultz, Sherri S.; Desbordes, Sabrina C.; Du, Zhuo; Kosiyatrakul, Settapong; Lipchina, Inna; Studer, Lorenz; Schildkraut, Carl L.

In: Molecular and Cellular Biology, Vol. 30, No. 18, 2010, p. 4521-4534.

Research output: Contribution to journalArticle

Schultz, Sherri S. ; Desbordes, Sabrina C. ; Du, Zhuo ; Kosiyatrakul, Settapong ; Lipchina, Inna ; Studer, Lorenz ; Schildkraut, Carl L. / Single-molecule analysis reveals changes in the DNA replication program for the POU5F1 locus upon human embryonic stem cell differentiation. In: Molecular and Cellular Biology. 2010 ; Vol. 30, No. 18. pp. 4521-4534.
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