Direct observation of individual tubulin dimers binding to growing microtubules

Keith J. Mickolajczyk, Elisabeth A. Geyer, Tae Kim, Luke M Rice, William O. Hancock

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

The biochemical basis of microtubule growth has remained elusive for over 30 years despite being fundamental for both cell division and associated chemotherapy strategies. Here, we combine interferometric scattering microscopy with recombinant tubulin to monitor individual tubulins binding to and dissociating from growing microtubule tips. We make direct, single-molecule measurements of tubulin association and dissociation rates. We detect two populations of transient dwell times and determine via binding-interface mutants that they are distinguished by the formation of one interprotofilament bond. Applying a computational model, we find that slow association kinetics with strong interactions along protofilaments best recapitulate our data and, furthermore, predicts plus-end tapering. Overall, we provide the most direct and complete experimental quantification of how microtubules grow to date.

Original languageEnglish (US)
Pages (from-to)7314-7322
Number of pages9
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number15
DOIs
StatePublished - Apr 9 2019

Keywords

  • Interferometric scattering microscopy
  • Microtubule dynamics
  • Single-molecule kinetics

ASJC Scopus subject areas

  • General

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