Cell cycle dependent phosphorylation and subnuclear organization of the histone gene regulator p220NPAT in human embryonic stem cells

Prachi N. Ghule, Klaus A. Becker, J. Wade Harper, Jane B. Lian, Janet L. Stein, Andre J. Van Wijnen, Gary S. Stein

Research output: Contribution to journalReview articlepeer-review

49 Scopus citations

Abstract

Human embryonic stem (ES) cells have an expedited cell cycle (∼15 h) due to an abbreviated G1 phase (∼2.5 h) relative to somatic cells. One principal regulatory event during cell cycle progression is the G1/S phase induction of histone biosynthesis to package newly replicated DNA. In somatic cells, histone H4 gene expression is controlled by CDK2 phosphorylation of p220NPAT and localization of HiNF-P/p220NPAT complexes with histone genes at Cajal body related subnuclear foci. Here we show that this 'S point' pathway is operative in situ in human ES cells (H9 cells; NIH-designated WA09). Immunofluorescence microscopy shows an increase in p220NPAT foci in G1 reflecting the assembly of histone gene regulatory complexes in situ. In contrast to somatic cells where duplication of p220NPAT foci is evident in S phase, the increase in the number of p220NPAT foci in ES cells appears to precede the onset of DNA synthesis as measured by BrdU incorporation. Phosphorylation of p220 NPAT at CDK dependent epitopes is most pronounced in S phase when cells exhibit elevated levels of cyclins E and A. Our data indicate that subnuclear organization of the HiNF-P/p220NPAT pathway is rapidly established as ES cells emerge from mitosis and that p220NPAT is subsequently phosphorylated in situ. Our findings establish that the HiNF-P/p220NPAT gene regulatory pathway operates in a cell cycle dependent microenvironment that supports expression of DNA replication-linked histone genes and chromatin assembly to accommodate human stem cell self-renewal.

Original languageEnglish (US)
Pages (from-to)9-17
Number of pages9
JournalJournal of Cellular Physiology
Volume213
Issue number1
DOIs
StatePublished - Oct 2007

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

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