An unbiased screen identifies DEP-1 tumor suppressor as a phosphatase controlling EGFR endocytosis.
BACKGROUND
The epidermal growth factor (EGF) stimulates rapid tyrosine phosphorylation of the EGF receptor (EGFR). This event precedes signaling from both the plasma membrane and from endosomes, and it is essential for recruitment of a ubiquitin ligase, CBL, that sorts activated receptors to endosomes and degradation. Because hyperphosphorylation of EGFR is involved in oncogenic pathways, we performed an unbiased screen of small interfering RNA (siRNA) oligonucleotides targeting all human tyrosine phosphatases.


RESULTS
We report the identification of PTPRK and PTPRJ (density-enhanced phosphatase-1 [DEP-1]) as EGFR-targeting phosphatases. DEP-1 is a tumor suppressor that dephosphorylates and thereby stabilizes EGFR by hampering its ability to associate with the CBL-GRB2 ubiquitin ligase complex. DEP-1 silencing enhanced tyrosine phosphorylation of endosomal EGFRs and, accordingly, increased cell proliferation. In line with functional interactions, EGFR and DEP-1 form physical associations, and EGFR phosphorylates a substrate-trapping mutant of DEP-1. Interestingly, the interactions of DEP-1 and EGFR are followed by physical segregation: whereas EGFR undergoes endocytosis, DEP-1 remains confined to the cell surface.


CONCLUSIONS
EGFR and DEP-1 physically interact at the cell surface and maintain bidirectional enzyme-substrate interactions, which are relevant to their respective oncogenic and tumor-suppressive functions. These observations highlight the emerging roles of vesicular trafficking in malignant processes.
Tarcic
Gabi
G
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel.
Boguslavsky
Shlomit K
SK
Wakim
Jean
J
Kiuchi
Tai
T
Liu
Angela
A
Reinitz
Felicia
F
Nathanson
David
D
Takahashi
Takamune
T
Mischel
Paul S
PS
Ng
Tony
T
Yarden
Yosef
Y
D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D066246:ErbB Receptors / Q000032:analysis / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D010766:Phosphorylation; D034622:RNA Interference; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3 / Q000032:analysis / Q000037:antagonists & inhibitors / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism
D004791:Enzyme Inhibitors; D044767:Ubiquitin-Protein Ligases; D066246:ErbB Receptors; C486288:PTPRJ protein, human; D054631:Receptor-Like Protein Tyrosine Phosphatases, Class 3