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bioRxiv · 10.1101/2023.05.03.539131

Interface-acting nucleotide controls polymerization dynamics at microtubule plus- and minus-ends

McCormick, L. A.·Cleary, J. M.·Hancock, W. O.·Rice, L. M.
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Abstract

GTP-tubulin is preferentially incorporated at growing microtubule ends, but the biochemical mechanism by which the bound nucleotide regulates the strength of tubulin:tubulin interactions is debated. The self-acting (cis) model posits that the nucleotide (GTP or GDP) bound to a particular tubulin dictates how strongly that tubulin interacts, whereas the interface-acting (trans) model posits that the nucleotide at the interface of two tubulin dimers is the determinant. We identified a testable difference between these mechanisms using mixed nucleotide simulations of microtubule elongation: with self-acting nucleotide, plus- and minus-end growth rates decreased in the same proportion to the amount of GDP-tubulin, whereas with interface-acting nucleotide, plus-end growth rates decreased disproportionately. We then experimentally measured plus- and minus-end elongation rates in mixed nucleotides and observed a disproportionate effect of GDP-tubulin on plus-end growth rates. Simulations of microtubule growth were consistent with GDP-tubulin binding at and poisoning plus-ends but not at minus-ends. Quantitative agreement between simulations and experiments required nucleotide exchange at terminal plus-end subunits to mitigate the poisoning effect of GDP-tubulin there. Our results indicate that the interfacial nucleotide determines tubulin:tubulin interaction strength, thereby settling a longstanding debate over the effect of nucleotide state on microtubule dynamics.

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McCormick, L. A., Cleary, J. M., Hancock, W. O., Rice, L. M.. 2023-05-04. Interface-acting nucleotide controls polymerization dynamics at microtubule plus- and minus-ends. https://doi.org/10.1101/2023.05.03.539131

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