Tesk1 interacts with Spry2 to abrogate its inhibition of ERK phosphorylation downstream of receptor tyrosine kinase signaling.
The Sprouty (Spry) proteins function as inhibitors of the Ras-ERK pathway downstream of various receptor tyrosine kinases. In this study, we have identified Tesk1 (testicular protein kinase 1) as a novel regulator of Spry2 function. Endogenous Tesk1 and Spry2 exist in a complex in cell lines and mouse tissues. Tesk1 coexpression relocalizes Spry2 to vesicles including endosomes, inhibiting its translocation to membrane ruffles upon growth factor stimulation. Independent of its kinase activity, Tesk1 binding leads to a loss of Spry2 function as an inhibitor of ERK phosphorylation and reverses inhibition of basic fibroblast growth factor (bFGF)- and nerve growth factor-induced neurite outgrowth in PC12 cells by Spry2. Furthermore, depletion of endogenous Tesk1 in PC12 cells leads to a reduction in neurite outgrowth induced by bFGF. Tesk1 nullifies the inhibitory effect of Spry2 by abrogating its interaction with the adaptor protein Grb2 and interfering with its serine dephosphorylation upon bFGF and FGF receptor 1 stimulation by impeding its binding to the catalytic subunit of protein phosphatase 2A. A construct of Tesk1 that binds to Spry2 but does not localize to the vesicles does not interfere with its function, highlighting the importance of subcellular localization of Tesk1 in this context. Conversely, Tesk1 does not affect interaction of Spry2 with the E3 ubiquitin ligase, c-Cbl, and consequently, does not affect its inhibition of Cbl-mediated ubiquitination of the epidermal growth factor receptor. By selectively modulating the downstream effects of Spry2, Tesk1 may thus serve as a molecular determinant of the signaling outcome.
Chandramouli
Sumana
S
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673.
Yu
Chye Yun
CY
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673.
Yusoff
Permeen
P
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673.
Lao
Dieu-Hung
DH
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673.
Leong
Hwei Fen
HF
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673.
Mizuno
Kensaku
K
Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.
Guy
Graeme R
GR
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673. Electronic address: mcbgg@imcb.a-star.edu.sg.
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D015536:Down-Regulation; D066246:ErbB Receptors / Q000378:metabolism; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D016501:Neurites / Q000378:metabolism; D016716:PC12 Cells; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D021381:Protein Transport; D050721:Proto-Oncogene Proteins c-cbl / Q000378:metabolism; D051381:Rats; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D015398:Signal Transduction; D013347:Subcellular Fractions; D054875:Ubiquitination; D018631:ras Proteins / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D012097:Repressor Proteins; C433763:Spred1 protein, mouse; C433764:Spred2 protein, mouse; D012694:Serine; D050721:Proto-Oncogene Proteins c-cbl; C097507:testis-specific protein kinase 1; D066246:ErbB Receptors; D020794:Receptor Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C118389:Spry2 protein, mouse; D048049:Extracellular Signal-Regulated MAP Kinases; D018631:ras Proteins