Abstract:
:The von Hippel-Lindau (VHL) protein protects microtubules (MTs) from destabilization by nocodazole treatment. Based on this fixed-cell assay with static end points, VHL has been reported to directly stabilize the MT cytoskeleton. To investigate the dynamic changes in MTs induced by VHL in living cells, we measured the influence of VHL on tubulin turnover using fluorescence recovery after photobleaching (FRAP). To this end, we engineered VHL-deficient renal cell carcinoma cells to constitutively incorporate fluorescently labeled tubulin and to inducibly express VHL. Induction of VHL in these cells resulted in a decrease of tubulin turnover as measured by FRAP at the cell periphery, while minimally influencing MT dynamics around the centrosome. Our data indicates that VHL changes the behavior of MTs dependent on their subcellular localization implying a role for VHL in cellular processes such as migration, polarization, and cell-cell interactions. Here we propose a complementary method to directly measure VHL-induced subcellular changes in microtubule dynamics, which may serve as a tool to study the effect of MT binding proteins such as VHL.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Lolkema MP,Mehra N,Jorna AS,van Beest M,Giles RH,Voest EEdoi
10.1016/j.yexcr.2004.07.016keywords:
subject
Has Abstractpub_date
2004-12-10 00:00:00pages
139-46issue
2eissn
0014-4827issn
1090-2422pii
S0014-4827(04)00394-5journal_volume
301pub_type
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