Oxidative signalling and gene expression during lymphocyte activation

  • Goldstone S
  • Milligan A
  • Hunt N
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Abstract

We previously have demonstrated an obligatory requirement for intracellular reactive oxygen species (ROS) generation during T lymphocyte activation, and have proposed that ROS may act as signalling agents in the regulation of certain cellular processes, for example, during cell cycle entry. In order to test this hypothesis, we have been interested to determine which, if any, cell cycle entry events are affected by oxidative signalling. Given the requirement for both oxidative signalling and altered gene expression during the G0to G1phase transition, we have attempted to establish the extent to which oxidative signalling affects global gene expression patterns during cell cycle entry, and to isolate and characterize mRNAs whose expression patterns are responsive to oxidative signalling during this process. Using differential display in a phenotypic screening approach, we have identified 10 mRNA species whose expression patterns were altered in response to inhibition of oxidative signalling during cell cycle entry. The expression patterns of 4 of these 10 mRNAs were unaffected during cell cycle arrest caused by a different mechanism, cyclosporin A-induced interference with calcineurin-mediated signalling events, implying that the altered expression patterns seen were not simply a consequence of cell cycle arrest. This suggests that the expression of these 4 mRNAs is regulated by a mechanism both necessary for cell cycle entry and sensitive to oxidative signalling RNAse protection assays confirmed that 2 of these 4 mRNAs were indeed responsive to redox regulation. These observations strongly suggest an involvement for oxidative signalling in the regulation of gene expression during the G0to G1phase transition, in peripheral blood mononuclear cells at least.

Author-supplied keywords

  • Cell cycle
  • Differential display
  • Gene expression
  • Lymphocyte activation
  • Oxidative signalling
  • Reactive oxygen species
  • Redox control

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Authors

  • Sherilyn D. Goldstone

  • Amanda D. Milligan

  • Nicholas H. Hunt

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