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Regulation of Set9-mediated H4K20 methylation by a PWWP domain protein

Yu Wang1, Bharat Reddy, James Thompson

  • 1Department of Biological Sciences, Columbia University, New York, NY 10027, USA.

Molecular Cell
|March 3, 2009
PubMed

Insights

A PWWP domain protein, Pdp1, is crucial for histone H4 lysine 20 (H4K20me) methylation by recruiting the Set9 enzyme to DNA. This epigenetic regulation is vital for DNA damage checkpoint activation.

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Chromatin Biology

Background:

  • Histone H4 lysine 20 methylation (H4K20me) is critical for DNA damage response.
  • The mechanisms regulating the H4K20 methyltransferase Set9 are not well understood.

Purpose of the Study:

  • To identify factors regulating Set9 function.
  • To investigate the role of Pdp1 in H4K20 methylation and DNA damage response.

Main Methods:

  • Protein-protein interaction studies to identify Set9-associated factors.
  • Chromatin immunoprecipitation to assess protein localization.
  • In vitro binding assays to characterize PWWP domain interactions.
  • Genotoxicity sensitivity assays.

Main Results:

  • Pdp1 was identified as a Set9-associated protein essential for Set9 chromatin localization.
  • Cells lacking Pdp1 showed defects in H4K20 methylation, genotoxic stress sensitivity, and Crb2 recruitment.
  • The PWWP domain of Pdp1 directly binds H4K20me, and this interaction is critical for Set9 chromatin association and H4K20 methylation in vivo.

Conclusions:

  • Pdp1 functions as a histone-binding protein that recruits Set9 to chromatin.
  • The PWWP domain acts as a novel methyl-lysine recognition motif, playing a key role in epigenetic regulation of DNA damage response.

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