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DNA-activated protein kinase functions in a newly observed S phase checkpoint that links histone mRNA abundance with

Berndt Müller1, Jane Blackburn, Carmen Feijoo

  • 1School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, Scotland, UK.

The Journal of Cell Biology
|December 26, 2007
PubMed

Insights

DNA replication and histone synthesis are linked. Inhibiting DNA-activated protein kinase (DNA-PK) with LY294002 disrupts this link, affecting histone mRNA levels during replication stress.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • DNA replication and histone synthesis are coupled processes essential for maintaining gene expression.
  • Histone gene expression relies on ongoing DNA replication.
  • Replication stress can lead to down-regulation of histone mRNA.

Purpose of the Study:

  • To investigate the molecular mechanisms coupling DNA replication and histone gene expression.
  • To determine the role of specific kinases in regulating histone mRNA abundance during replication stress.
  • To identify the target of LY294002 in uncoupling DNA replication from histone mRNA levels.

Main Methods:

  • Treatment of S phase-arrested cells with kinase inhibitors caffeine and LY294002.
  • Analysis of histone mRNA abundance and DNA replication.
  • Investigating the involvement of ATR/ATM and DNA-PK signaling pathways.
  • Assessing the phosphorylation of UPF1 and other proteins.

Main Results:

  • Caffeine and LY294002 uncouple DNA replication from histone mRNA abundance, altering replication stress-induced histone mRNA down-regulation.
  • ATR/ATM kinases are not solely responsible for coupling DNA replication and histone mRNA levels.
  • LY294002's effect is dependent on functional DNA-PK, identifying it as the target.
  • Replication stress induces UPF1 phosphorylation, suggesting its role in histone mRNA stability control.

Conclusions:

  • DNA-PK plays a critical role in coupling DNA replication and histone mRNA stability.
  • Histone mRNA stability may be directly regulated by ATR/ATM and DNA-PK-mediated phosphorylation of UPF1.
  • Understanding this coupling mechanism provides insights into cellular response to replication stress.

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