TY - JOUR
T1 - Quantitative image analysis identifies pVHL as a key regulator of microtubule dynamic instability
AU - Thoma, Claudio R.
AU - Matov, Alexandre
AU - Gutbrodt, Katrin L.
AU - Hoerner, Christian R.
AU - Smole, Zlatko
AU - Krek, Wilhelm
AU - Danuser, Gaudenz
PY - 2010/9/20
Y1 - 2010/9/20
N2 - Von Hippel-Lindau (VHL) tumor suppressor gene mutations predispose carriers to kidney cancer. The protein pVHL has been shown to interact with microtubules (MTs), which is critical to cilia maintenance and mitotic spindle orientation. However, the function for pVHL in the regulation of MT dynamics is unknown. We tracked MT growth via the plus end marker EB3 (end-binding protein 3)-GFP and inferred additional parameters of MT dynamics indirectly by spatiotemporal grouping of growth tracks from live cell imaging. Our data establish pVHL as a near-optimal MT-stabilizing protein: it attenuates tubulin turnover, both during MT growth and shrinkage, inhibits catastrophe, and enhances rescue frequencies. These functions are mediated, in part, by inhibition of tubulin guanosine triphosphatase activity in vitro and at MT plus ends and along the MT lattice in vivo. Mutants connected to the VHL cancer syndrome are differentially compromised in these activities. Thus, single cell-level analysis of pVHL MT regulatory function allows new predictions for genotype to phenotype associations that deviate from the coarser clinically defined mutant classifications.
AB - Von Hippel-Lindau (VHL) tumor suppressor gene mutations predispose carriers to kidney cancer. The protein pVHL has been shown to interact with microtubules (MTs), which is critical to cilia maintenance and mitotic spindle orientation. However, the function for pVHL in the regulation of MT dynamics is unknown. We tracked MT growth via the plus end marker EB3 (end-binding protein 3)-GFP and inferred additional parameters of MT dynamics indirectly by spatiotemporal grouping of growth tracks from live cell imaging. Our data establish pVHL as a near-optimal MT-stabilizing protein: it attenuates tubulin turnover, both during MT growth and shrinkage, inhibits catastrophe, and enhances rescue frequencies. These functions are mediated, in part, by inhibition of tubulin guanosine triphosphatase activity in vitro and at MT plus ends and along the MT lattice in vivo. Mutants connected to the VHL cancer syndrome are differentially compromised in these activities. Thus, single cell-level analysis of pVHL MT regulatory function allows new predictions for genotype to phenotype associations that deviate from the coarser clinically defined mutant classifications.
UR - http://www.scopus.com/inward/record.url?scp=77957198722&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77957198722&partnerID=8YFLogxK
U2 - 10.1083/jcb.201006059
DO - 10.1083/jcb.201006059
M3 - Article
C2 - 20855504
AN - SCOPUS:77957198722
SN - 0021-9525
VL - 190
SP - 991
EP - 1003
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 6
ER -