Paxillin-dependent stimulation of microtubule catastrophes at focal adhesion sites.
An organized microtubule array is essential for the polarized motility of fibroblasts. Dynamic microtubules closely interact with focal adhesion sites in migrating cells. Here, we examined the effect of focal adhesions on microtubule dynamics. We observed that the probability of microtubule catastrophes (transitions from growth to shrinkage) was seven times higher at focal adhesions than elsewhere. Analysis of the dependence between the microtubule growth rate and catastrophe probability throughout the cytoplasm revealed that a nonspecific (mechanical or spatial) factor provided a minor contribution to the catastrophe induction by decreasing microtubule growth rate at adhesions. Strikingly, at the same growth rate, the probability of catastrophes was significantly higher at adhesions than elsewhere, indicative of a site-specific biochemical trigger. The observed catastrophe induction occurred at adhesion domains containing the scaffolding protein paxillin that has been shown previously to interact with tubulin. Furthermore, replacement of full-length paxillin at adhesion sites by microinjected paxillin LIM2-LIM3 domains suppressed microtubule catastrophes exclusively at adhesions. We suggest that paxillin influences microtubule dynamics at focal adhesions by serving as a scaffold for a putative catastrophe factor and/or regulating its exposure to microtubules.
Efimov
Andrey
A
Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN, USA.
Schiefermeier
Natalia
N
Grigoriev
Ilya
I
Ohi
Ryoma
R
Brown
Michael C
MC
Turner
Christopher E
CE
Small
J Victor
JV
Kaverina
Irina
I
D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D003593:Cytoplasm / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000378:metabolism; D006054:Goldfish; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D008870:Microtubules / Q000378:metabolism*; D008954:Models, Biological; D051419:Paxillin / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D003598:Cytoskeletal Proteins; D051419:Paxillin; D005982:Glutathione Transferase