Mobility of the von Hippel-Lindau tumour suppressor protein is regulated by kinesin-2.
The von Hippel-Lindau tumour suppressor protein (pVHL) participates in many cellular processes including oxygen sensing, microtubule stability and primary cilia regulation. Recently, we identified ATP-dependent motor complex kinesin-2 to endogenously bind the full-length variant of VHL (pVHL30) in primary kidney cells, and mediate its association to microtubules. Here we show that pVHL also endogenously binds the neuronal kinesin-2 complex, which slightly differs from renal kinesin-2. To investigate the role of kinesin-2 in pVHL mobility, we performed fluorescence recovery after photobleaching (FRAP) experiments in neuroblastoma cells. We observe that pVHL30 is a highly mobile cytoplasmic protein, which becomes an immobile centrosomal protein after ATP-depletion in living cells. This response to ATP-depletion is independent of GSK3beta-dependent phosphorylation of pVHL30. Furthermore, VHL variant alleles with reduced binding to kinesin-2 fail to respond to ATP-depletion. Accordingly, interfering with pVHL30-KIF3A interaction by either overexpressing a dominant negative construct or by reducing endogenous cellular levels of KIF3A by RNAi abolishes pVHL's response to ATP-depletion. From these data we suggest that mobility of a subcellular pool of pVHL is regulated by the ATP-dependent kinesin-2 motor. Kinesin-2 driven mobility of cytoplasmic pVHL might enable pVHL to function as a tumour suppressor.
Mans
Dorus A
DA
Laboratory of Experimental Oncology, Department of Medical Oncology, University Medical Center Utrecht, Utrecht, The Netherlands.
Lolkema
Martijn P
MP
van Beest
Moniek
M
Daenen
Laura G
LG
Voest
Emile E
EE
Giles
Rachel H
RH
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000483:Alleles; D000818:Animals; D002457:Cell Extracts; D002460:Cell Line; D036681:Fluorescence Recovery After Photobleaching; D006801:Humans; D047468:Immunoprecipitation; D016547:Kinesins / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014162:Transfection; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000378:metabolism*
D002457:Cell Extracts; C490202:Kif3a protein, mouse; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D016547:Kinesins