An important role for CDK2 in G1 to S checkpoint activation and DNA damage response in human embryonic stem cells

Irina Neganova, Felipe Vilella, Stuart P. Atkinson, Maria Lloret, Joao Passos, Thomas Von Zglinicki, José Enrique O'Connor, Deborah Burks, Richard Jones, Lyle Armstrong, Majlinda Lako

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

A precise understanding of mechanisms used by human embryonic stem cells (hESCs) to maintain genomic integrity is very important for their potential clinical applications. The G1 checkpoint serves to protect genomic integrity and prevents cells with damaged DNA from entering S-phase. Previously, we have shown that downregulation of cyclindependent kinase 2 (CDK2) in hESC causes G1 arrest, loss of pluripotency, upregulation of cell cycle inhibitors p21 and p27 and differentiation toward extraembryonic lineages. In this study, we investigate in detail the role of CDK2 in cellular processes, which are crucial to the maintenance of genomic stability in hESC such as G1 checkpoint activation, DNA repair, and apoptosis. Our results suggest that downregulation of CDK2 triggers the G1 checkpoint through the activation of the ATM-CHK2-p53-p21 pathway. Downregulation of CDK2 is able to induce sustained DNA damage and to elicit the DNA damage response (DDR) as evidenced by the formation of distinct c-H2.AX and RAD52-BRCA1 foci in hESC nuclei. CDK2 downregulation causes high apoptosis at the early time points; however, this is gradually decreased overtime as the DDR is initiated. Our mass spectrometry analysis suggest that CDK2 does interact with a large number of proteins that are involved in key cellular processes such as DNA replication, cell cycle progression, DNA repair, chromatin modeling, thus, suggesting a crucial role for CDK2 in orchestrating a fine balance between cellular proliferation, cell death, and DNA repair in hESC.

Original languageEnglish (US)
Pages (from-to)651-659
Number of pages9
JournalStem Cells
Volume29
Issue number4
DOIs
StatePublished - Apr 1 2011
Externally publishedYes

Fingerprint

DNA Damage
Phosphotransferases
Down-Regulation
DNA Repair
Cell Cycle
Apoptosis
Chromatin Assembly and Disassembly
Genomic Instability
Human Embryonic Stem Cells
Cell Nucleus
DNA Replication
S Phase
Mass Spectrometry
Cell Death
Up-Regulation
Maintenance
Cell Proliferation
DNA
Proteins

Keywords

  • Apoptosis
  • Ataxia telangiectasia mutated
  • CHK2
  • Cyclin-dependent kinase 2
  • DNA damage
  • G1 checkpoint
  • Human embryonic stem cells

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Neganova, I., Vilella, F., Atkinson, S. P., Lloret, M., Passos, J., Von Zglinicki, T., ... Lako, M. (2011). An important role for CDK2 in G1 to S checkpoint activation and DNA damage response in human embryonic stem cells. Stem Cells, 29(4), 651-659. https://doi.org/10.1002/stem.620

An important role for CDK2 in G1 to S checkpoint activation and DNA damage response in human embryonic stem cells. / Neganova, Irina; Vilella, Felipe; Atkinson, Stuart P.; Lloret, Maria; Passos, Joao; Von Zglinicki, Thomas; O'Connor, José Enrique; Burks, Deborah; Jones, Richard; Armstrong, Lyle; Lako, Majlinda.

In: Stem Cells, Vol. 29, No. 4, 01.04.2011, p. 651-659.

Research output: Contribution to journalArticle

Neganova, I, Vilella, F, Atkinson, SP, Lloret, M, Passos, J, Von Zglinicki, T, O'Connor, JE, Burks, D, Jones, R, Armstrong, L & Lako, M 2011, 'An important role for CDK2 in G1 to S checkpoint activation and DNA damage response in human embryonic stem cells', Stem Cells, vol. 29, no. 4, pp. 651-659. https://doi.org/10.1002/stem.620
Neganova, Irina ; Vilella, Felipe ; Atkinson, Stuart P. ; Lloret, Maria ; Passos, Joao ; Von Zglinicki, Thomas ; O'Connor, José Enrique ; Burks, Deborah ; Jones, Richard ; Armstrong, Lyle ; Lako, Majlinda. / An important role for CDK2 in G1 to S checkpoint activation and DNA damage response in human embryonic stem cells. In: Stem Cells. 2011 ; Vol. 29, No. 4. pp. 651-659.
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