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Quantification of microtubule stutters: dynamic instability behaviors that are strongly associated with catastrophe


Mahserejian SMShant M , Scripture JPJared P , Mauro AJAva J , Lawrence EJElizabeth J , Jonasson EMErin M , Murray KSKristopher S , Li JJun , Gardner MMelissa , Alber MMark , Zanic MMarija , Goodson HVHolly V . Molecular biology of the cell. 2022 2 2; 33(3). ar22


Microtubules (MTs) are cytoskeletal fibers that undergo dynamic instability (DI), a remarkable process involving phases of growth and shortening separated by stochastic transitions called catastrophe and rescue. Dissecting DI mechanism(s) requires first characterizing and quantifying these dynamics, a subjective process that often ignores complexity in MT behavior. We present a tatistical ool for utomated ynamic nstability nalysis (STADIA) that identifies and quantifies not only growth and shortening, but also a category of intermediate behaviors that we term “stutters.” During stutters, the rate of MT length change tends to be smaller in magnitude than during typical growth or shortening phases. Quantifying stutters and other behaviors with STADIA demonstrates that stutters precede most catastrophes in our in vitro experiments and dimer-scale MT simulations, suggesting that stutters are mechanistically involved in catastrophes. Related to this idea, we show that the anticatastrophe factor CLASP2γ works by promoting the return of stuttering MTs to growth. STADIA enables more comprehensive and data-driven analysis of MT dynamics compared with previous methods. The treatment of stutters as distinct and quantifiable DI behaviors provides new opportunities for analyzing mechanisms of MT dynamics and their regulation by binding proteins.