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Involvement of H4(D10S170) protein in ATM-dependent response to DNA damage

F Merolla1, F Pentimalli, R Pacelli

  • 1Dipartimento di Biologia e Patologia Cellulare e Molecolare, University 'Federico II', Naples, Italy.

Oncogene
|April 11, 2007
PubMed

Insights

The H4(D10S170) gene is phosphorylated by ATM kinase in response to DNA damage, influencing cell survival and apoptosis. Its dysfunction may contribute to thyroid cancer development.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Cancer Research

Background:

  • The H4(D10S170) gene is frequently rearranged with RET in papillary thyroid tumors (RET/PTC1).
  • The ataxia telangectasia mutated (ATM) kinase is a key regulator of DNA damage response pathways.
  • The role of H4(D10S170) in DNA damage signaling remained largely uncharacterized.

Purpose of the Study:

  • To investigate the involvement of H4(D10S170) in DNA damage response pathways.
  • To elucidate the relationship between H4(D10S170) phosphorylation and ATM kinase activity.
  • To determine the functional consequences of H4(D10S170) modulation on cellular response to genotoxic stress.

Main Methods:

  • Cell treatment with etoposide or ionizing radiation (IR).
  • Analysis of H4(D10S170) phosphorylation and localization in various cell lines, including ATM-deficient cells.
  • Assessment of apoptosis, DNA synthesis, and mitotic progression following H4(D10S170) manipulation (silencing or mutation).
  • Clonogenic assays and bromodeoxyuridine incorporation assays.

Main Results:

  • H4(D10S170) undergoes ATM-mediated phosphorylation at Thr 434 in response to DNA damage, stabilizing its nuclear localization.
  • In ATM-deficient cells, H4(D10S170) is cytoplasmic and not phosphorylated after IR exposure.
  • Inhibition of ATM kinase impairs H4(D10S170) apoptotic activity.
  • H4(T434A) mutant expression protects cells from genotoxic stress-induced apoptosis.
  • Silencing H4(D10S170) enhances cell survival, DNA synthesis, and mitotic progression after IR exposure.

Conclusions:

  • H4(D10S170) is a downstream target of ATM-mediated DNA damage signaling.
  • Phosphorylation of H4(D10S170) by ATM is crucial for its role in apoptosis and cellular response to DNA damage.
  • Dysregulation of H4(D10S170) function may play a role in thyroid carcinogenesis.

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