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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Spry2 does not directly modulate Raf-1 kinase activity in v-Ha-ras-transformed NIH 3T3 fibroblasts

  • Ahn, Jun-Ho (Department of Biology, College of Natural Sciences, University of Incheon) ;
  • Eum, Ki-Hwan (Department of Biology, College of Natural Sciences, University of Incheon) ;
  • Lee, Michael (Department of Biology, College of Natural Sciences, University of Incheon)
  • Published : 2010.03.31
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Abstract

Sprouty (Spry) proteins have previously been suggested as negative regulators of the MAPK pathway through interaction with Raf-1. However, the molecular basis of this inhibition has not been elucidated. In this study, we used cells expressing FLAGtagged Raf-1 with point mutations at known phosphorylation sites to reveal that activation of Raf-1 mutants does not correlate with their degree of interaction with Spry2. The association of Raf-1 with Spry2 in intact cells was further corroborated by immunofluorescence colocalization. Additionally, there was no significant change observed in the strength of interaction between Raf-1 mutants and Spry2 after paclitaxel treatment despite differences in the activation levels of these mutants. Thus, our study provides the evidence that Spry2 does not directly regulate Raf-1 kinase activity, but instead acts as a scaffolding protein that assists interactions between Raf-1 kinase and its direct regulators.

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References

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