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Current Genetics

, Volume 46, Issue 2, pp 72–81 | Cite as

The yeast phosphotyrosyl phosphatase activator protein, yPtpa1/Rrd1, interacts with Sit4 phosphatase to mediate resistance to 4-nitroquinoline-1-oxide and UVA

  • J. Douville
  • J. David
  • P.-K. Fortier
  • Dindial RamotarEmail author
Research Article

Abstract

We previously reported the isolation of mutants hypersensitive to the genotoxic agent 4-nitroquinoline-1-oxide, a potent inducer of oxidative stress. One of the mutants was defective in a gene designated yPTPA1, encoding a protein related to the human phosphotyrosyl phosphatase activator hPTPA, which is believed to play a role in activating the serine/threonine phosphatase PP2A. Yeast yptpa1Δ mutants are also sensitive to the UVA component of sunlight known to produce reactive oxygen species, suggesting a role for yPtpa1 in oxidative stress response. We now report the characterization of another 4-nitroquinoline-1-oxide-sensitive mutant, EBY20. We show that this mutant is defective in the SIT4 gene encoding a catalytic subunit of the PP2A phosphatases and that sit4Δ mutants exhibit hypersensitivity to 4-nitroquinoline-1-oxide and UVA, but not to UVC at 254 nm. Like the yptpa1Δ mutants, sit4Δ mutants are also defective in the repair of 4-nitroquinoline-1-oxide-induced DNA lesions. Genetic analysis revealed that both yPtpa1 and Sit4 function in the same pathway to protect cells against the lethal effects of 4-nitroquinoline-1-oxide and UVA. Moreover, we demonstrate that yPtpa1-affinity columns specifically retain Sit4, confirming a previous report that these two proteins indeed belong to a complex. Cellular localization studies using GFP-tagged proteins reveals that yPtpa1 is localized to the cytoplasm and the nucleus, while the Sit4 protein shows an intense staining spot in the cytoplasm and diffused staining in this organelle. We suggest that the yPtpa1–Sit4 complex may participate in a novel mechanism that mediates repair of oxidative DNA damage caused by 4-nitroquinoline-1-oxide and UVA.

Keywords

Parent Strain SIT4 Gene Methyl Methane Sulfonate PP2A Phosphatase Variable Subunit 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

We thank Anick Leduc for assistance with the microscope and Cyril Sabbah for reading the manuscript. This study was supported by a grant to D.R. from the Canadian Institutes of Health Research. During the tenure of this work, D.R. was supported by a career scientist award from the NCIC, and currently by a senior fellowship from the Fonds de la Recherche en Sante du Quebec.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • J. Douville
    • 1
  • J. David
    • 1
  • P.-K. Fortier
    • 1
  • Dindial Ramotar
    • 1
    Email author
  1. 1.Centre de Recherche, Hopital Maisonneuve-RosemontUniversité de MontréalMontréalCanada

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