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Limited nucleotide pools restrict Epstein-Barr virus-mediated B-cell immortalization.

A Y Hafez1, J E Messinger1, K McFadden1

  • 1Department of Molecular Genetics and Microbiology, Center for Virology, Duke University School of Medicine, Durham, NC, USA.

Oncogenesis
|June 13, 2017
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Summary

Epstein-Barr virus (EBV) infection causes B-cell hyper-proliferation, but limited purine deoxyribonucleotide triphosphate (dNTP) pools cause DNA damage. Supplementing nucleosides enhances EBV B-cell immortalization by reducing replicative stress.

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Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Epstein-Barr virus (EBV) infection of B cells causes hyper-proliferation.
  • Many EBV-infected cells undergo senescence due to ATM/Chk2-mediated growth arrest.
  • DNA damage response is activated by oncogenes and tumor viruses.

Purpose of the Study:

  • To investigate the role of DNA replicative stress and nucleotide pools in limiting EBV-infected B-cell outgrowth.
  • To understand the mechanisms behind EBV-mediated B-cell immortalization.

Main Methods:

  • Assessed the ataxia telangiectasia and Rad3-related (ATR) signaling pathway activation.
  • Performed nuclear halo assays to measure replicative stress and DNA damage.
  • Quantified deoxyribonucleotide triphosphate (dNTP) pools in EBV-infected cells.
  • Supplemented cells with exogenous nucleosides.

Main Results:

  • EBV triggered ATR pathway activation in early proliferating cells, increasing sensitivity to ATR inhibition.
  • Early EBV-infected cells showed higher replicative stress and DNA damage than late proliferating cells.
  • Early hyper-proliferating B cells had limited purine dNTP pools compared to later stages.
  • Exogenous nucleoside supplementation enhanced B-cell immortalization and rescued replicative stress.

Conclusions:

  • Purine deoxyribonucleotide triphosphate (dNTP) biosynthesis is critical for early EBV-mediated B-cell immortalization.
  • Replicative stress and limited dNTP pools restrict EBV-driven B-cell proliferation.
  • Targeting dNTP metabolism could be a strategy to modulate EBV-induced B-cell immortalization.