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. 2013 May 15;18(2):297–314. doi: 10.2478/s11658-013-0090-3

PTPN4 negatively regulates CrkI in human cell lines

Juan Zhou 190, Bingbing Wan 190, Jingxuan Shan 190, Huili Shi 190, Yanhong Li 190,✉, Keke Huo 190,✉
PMCID: PMC6275623  PMID: 23666597

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.

Electronic Supplementary Material

Supplementary material is available for this article at 10.2478/s11658-013-0090-3 and is accessible for authorized users.

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

Full Text

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Abbreviations used

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

Contributor Information

Yanhong Li, Phone: +86-21-65642339, Email: lyhong@fudan.edu.cn.

Keke Huo, Phone: +86-21-55664526, FAX: +86-21-55664526, Email: kkhuo@fudan.edu.cn.

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