A role for NANOG in G1 to S transition in human embryonic stem cells through direct binding of CDK6 and CDC25A.

The Journal of Cell Biology
X ZhangMajlinda Lako

Abstract

In this study, we show that NANOG, a master transcription factor, regulates S-phase entry in human embryonic stem cells (hESCs) via transcriptional regulation of cell cycle regulatory components. Chromatin immunoprecipitation combined with reporter-based transfection assays show that the C-terminal region of NANOG binds to the regulatory regions of CDK6 and CDC25A genes under normal physiological conditions. Decreased CDK6 and CDC25A expression in hESCs suggest that both CDK6 and CDC25A are involved in S-phase regulation. The effects of NANOG overexpression on S-phase regulation are mitigated by the down-regulation of CDK6 or CDC25A alone. Overexpression of CDK6 or CDC25A alone can rescue the impact of NANOG down-regulation on S-phase entry, suggesting that CDK6 and CDC25A are downstream cell cycle effectors of NANOG during the G1 to S transition.

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Citations

Aug 8, 2009·Cellular and Molecular Life Sciences : CMLS·Manal BosnaliFrank Edenhofer
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Methods Mentioned

BETA
transfection
flow cytometry
ChIP
immunoprecipitation
PCR
Flow
transfections
FCS
immunoprecipitation assay
Assay

Software Mentioned

AxioVision
ModFit
LAS
CellQuest
3000
LightCycler relative quantification
FlowJo
GeneSpring
LightCycler

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