PAK4 kinase activity and somatic mutation promote carcinoma cell motility and influence inhibitor sensitivity.
Hepatocyte growth factor (HGF) and its receptor (c-Met) are associated with cancer cell motility and invasiveness. p21-activated kinase 4 (PAK4), a potential therapeutic target, is recruited to and activated by c-Met. In response, PAK4 phosphorylates LIM kinase 1 (LIMK1) in an HGF-dependent manner in metastatic prostate carcinoma cells. PAK4 overexpression is known to induce increased cell migration speed but the requirement for kinase activity has not been established. We have used a panel of PAK4 truncations and mutations in a combination of overexpression and RNAi rescue experiments to determine the requirement for PAK4 kinase activity during carcinoma cell motility downstream of HGF. We find that neither the kinase domain alone nor a PAK4 mutant unable to bind Cdc42 is able to fully rescue cell motility in a PAK4-deficient background. Nevertheless, we find that PAK4 kinase activity and associated LIMK1 activity are essential for carcinoma cell motility, highlighting PAK4 as a potential anti-metastatic therapeutic target. We also show here that overexpression of PAK4 harbouring a somatic mutation, E329K, increased the HGF-driven motility of metastatic prostate carcinoma cells. E329 lies within the glycine-rich loop region of the kinase. Our data suggest that E329K mutation leads to a modest increase in kinase activity, conferring resistance to competitive ATP inhibitors in addition to promoting cell migration. The existence of such a mutation may have implications for the development of PAK4-specific competitive ATP inhibitors should PAK4 be further explored for clinical inhibition.
Randall Division of Cell and Molecular Biophysics, King's College London, London, UK.
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D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D054461:Lim Kinases / Q000378:metabolism; D008297:Male; D009154:Mutation; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D015398:Signal Transduction; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D054462:p21-Activated Kinases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; C098587:GAB1 protein, human; D017228:Hepatocyte Growth Factor; D000255:Adenosine Triphosphate; C116367:PAK4 protein, human; C515892:LIMK1 protein, human; D054461:Lim Kinases; D054462:p21-Activated Kinases; D020764:cdc42 GTP-Binding Protein