Neuronal axons terminate as synaptic boutons that form stable yet plastic connections with their targets. Synaptic bouton development relies on an underlying network of both long-lived and dynamic microtubules that provide structural stability for the boutons while also allowing for their growth and remodeling. However, a molecular-scale mechanism that explains how neurons appropriately balance these two microtubule populations remains a mystery. We hypothesized that α-tubulin acetyltransferase (αTAT), which both stabilizes long-lived microtubules against mechanical stress via acetylation and has been implicated in promoting microtubule dynamics, could play a role in this process. Using the Drosophila neuromuscular junction as a model, we found that non-enzymatic dαTAT activity limits the growth of synaptic boutons by affecting dynamic, but not stable, microtubules. Loss of dαTAT results in the formation of ectopic boutons. These ectopic boutons can be similarly suppressed by resupplying enzyme-inactive dαTAT or by treatment with a low concentration of the microtubule-targeting agent vinblastine, which acts to suppress microtubule dynamics. Biophysical reconstitution experiments revealed that non-enzymatic αTAT1 activity destabilizes dynamic microtubules but does not substantially impact the stability of long-lived microtubules. Further, during microtubule growth, non-enzymatic αTAT1 activity results in increasingly extended tip structures, consistent with an increased rate of acceleration of catastrophe frequency with microtubule age, perhaps via tip structure remodeling. Through these mechanisms, αTAT enriches for stable microtubules at the expense of dynamic ones. We propose that the specific suppression of dynamic microtubules by non-enzymatic αTAT activity regulates the remodeling of microtubule networks during synaptic bouton development. Blending in vivo and in vitro reconstitution approaches, Coombes and Saunders et al. reveal that α-tubulin acetyltransferase uses its non-enzymatic activity to curb synaptic terminal growth in neurons. Non-enzymatic αTAT activity accelerates microtubule tip structure aging, which selectively destabilizes dynamic, but not stable, microtubules.
Bibliographical noteFunding Information:
This work was supported by grant R21NS101553 from the NINDS of the National Institutes of Health to J.W. and M.K.G. Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the NSF -funded Materials Research Facilities Network ( https://www.mrfn.org ) via the MRSEC program. We thank members of the Wildonger and Gardner laboratories for helpful discussions.
- microtubule aging
- neuromuscular junction
- synaptic bouton
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural