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Paxillin-dependent stimulation of microtubule catastrophes at focal adhesion sites
Andrey Efimov1, Natalia Schiefermeier, Ilya Grigoriev
1Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN, USA.
Abstract:
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.
Insights
Focal adhesions increase microtubule catastrophes sevenfold via a biochemical trigger involving paxillin. This suggests paxillin acts as a scaffold, influencing microtubule dynamics at these crucial cellular sites.
Area of Science:
- Cell Biology
- Cytoskeletal Dynamics
- Cell Migration
Background:
- Microtubule organization is vital for fibroblast motility.
- Dynamic microtubules interact with focal adhesions during cell migration.
Purpose of the Study:
- To investigate the impact of focal adhesions on microtubule dynamics.
- To elucidate the molecular mechanisms underlying microtubule behavior at focal adhesions.
Main Methods:
- Quantitative analysis of microtubule growth and catastrophe events.
- Microinjection of paxillin domains to assess functional roles.
- Localization studies of paxillin and microtubules at focal adhesions.
Main Results:
- Microtubule catastrophe probability was seven times higher at focal adhesions.
- A site-specific biochemical trigger, not just mechanical factors, drives catastrophes at adhesions.
- Paxillin at adhesion sites correlated with increased microtubule catastrophes.
- Specific paxillin domains (LIM2-LIM3) suppressed adhesion-specific catastrophes.
Conclusions:
- Paxillin plays a key role in regulating microtubule dynamics at focal adhesions.
- Paxillin may act as a scaffold for factors inducing microtubule catastrophes.
- This interaction influences cytoskeletal organization and cell migration.
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