A crucial role for DOK1 in PDGF-BB-stimulated glioma cell invasion through p130Cas and Rap1 signalling.
DOK1 regulates platelet-derived growth factor (PDGF)-BB-stimulated glioma cell motility. Mechanisms regulating tumour cell motility are essential for invasion and metastasis. We report here that PDGF-BB-mediated glioma cell invasion and migration are dependent on the adaptor protein downstream of kinase 1 (DOK1). DOK1 is expressed in several glioma cell lines and in tumour biopsies from high-grade gliomas. DOK1 becomes tyrosine phosphorylated upon PDGF-BB stimulation of human glioma cells. Knockdown of DOK1 or expression of a DOK1 mutant (DOK1FF) containing Phe in place of Tyr at residues 362 and 398, resulted in inhibition of both the PDGF-BB-induced tyrosine phosphorylation of p130Cas (also known as BCAR1) and the activation of Rap1. DOK1 colocalises with tyrosine phosphorylated p130Cas at the cell membrane of PDGF-BB-treated cells. Expression of a non-tyrosine-phosphorylatable substrate domain mutant of p130Cas (p130Cas15F) inhibited PDGF-BB-mediated Rap1 activation. Knockdown of DOK1 and Rap1 inhibited PDGF-BB-induced chemotactic cell migration, and knockdown of DOK1 and Rap1 and expression of DOK1FF inhibited PDGF-mediated three-dimensional (3D) spheroid invasion. These data show a crucial role for DOK1 in the regulation of PDGF-BB-mediated tumour cell motility through a p130Cas-Rap1 signalling pathway. [Corrected]
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Angela
A
Barrett
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Ian M
IM
Evans
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Antonina
A
Frolov
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Gary
G
Britton
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Caroline
C
Pellet-Many
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Maiko
M
Yamaji
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Vedanta
V
Mehta
Reta Lila Weston Institute of Neurological Studies.
Rina
R
Bandopadhyay
Division of Neuropathology and Department of Neurodegenerative Disease, Institute of NeurologyUniversity College LondonLondon WC1E 6JJUK.
Ningning
N
Li
Division of Neuropathology and Department of Neurodegenerative Disease, Institute of NeurologyUniversity College LondonLondon WC1E 6JJUK.
Sebastian
S
Brandner
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK.
Ian C
IC
Zachary
Centre for Cardiovascular Biology and Medicine, Division of Medicine, Rayne Building, University College London, London WC1E 6JJ, UK p.frankel@ucl.ac.uk.
Paul
P
Frankel
D000077214:Becaplermin; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002633:Chemotaxis; D050719:Crk-Associated Substrate Protein / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology*; D005909:Glioblastoma / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D009361:Neoplasm Invasiveness; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010750:Phosphoproteins / Q000502:physiology*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D020574:Proto-Oncogene Proteins c-sis / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000502:physiology*; D000089804:Shelterin Complex; D015398:Signal Transduction; D034501:Telomere-Binding Proteins / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism
D000077214:Becaplermin; D050719:Crk-Associated Substrate Protein; D004268:DNA-Binding Proteins; D019869:Phosphatidylinositol 3-Kinases; D010750:Phosphoproteins; D020574:Proto-Oncogene Proteins c-sis; D016601:RNA-Binding Proteins; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins; D019061:src-Family Kinases; C493841:BCAR1 protein, human; C489560:DOK1 protein, human; C440809:TERF2IP protein, human
Cell motility; DOK1; GTPase; PDGF signalling; p130Cas