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PTPN4 negatively regulates CrkI in human cell lines

Abstract

PTPN4 is a widely expressed non-receptor protein tyrosine phosphatase. Although its overexpression inhibits cell growth, the proteins with which it interacts to regulate cell growth are unknown. In this study, we identified CrkI as a PTPN4-interacting protein using a yeast two-hybrid, and confirmed this interaction using in vitro GST pull-down and co-immunoprecipitation and co-localization assays. We further determined the interactional regions as the SH3 domain of CrkI and the proline-rich region between amino acids 462 and 468 of PTPN4. Notably, overexpression of PTPN4 inhibits CrkI-mediated proliferation and wound healing of HEK293T cells, while knockdown of PTPN4 by siRNA in Hep3B cells enhances CrkI-mediated cell growth and motility. Moreover, our data show that ectopic expression of PTPN4 reduces the phosphorylation level of CrkI in HEK293T cells. These findings suggest that PTPN4 negatively regulates cell proliferation and motility through dephosphorylation of CrkI.

Abbreviations

DAPI:

4′,6-diamidino-2-phenylindole

ORF:

open-reading frame

PTPs:

protein tyrosine phosphatases

RNAi:

RNA interference

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Correspondence to Yanhong Li or Keke Huo.

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Zhou, J., Wan, B., Shan, J. et al. PTPN4 negatively regulates CrkI in human cell lines. Cell Mol Biol Lett 18, 297–314 (2013). https://doi.org/10.2478/s11658-013-0090-3

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  • DOI: https://doi.org/10.2478/s11658-013-0090-3

Key words

  • PTPN4
  • PTPMEG
  • CrkI
  • CRK
  • Cell proliferation
  • Protein tyrosine phosphatase
  • Wound-healing assay
  • Subcellular localization
  • Protein-protein interactions
  • Yeast two-hybrid