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

6 Citations (Scopus)

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

Fingerprint

Tubulin
Microtubules
Observation
Cell Division
Microscopy
Drug Therapy
Growth
Population

Keywords

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

ASJC Scopus subject areas

  • General

Cite this

Direct observation of individual tubulin dimers binding to growing microtubules. / Mickolajczyk, Keith J.; Geyer, Elisabeth A.; Kim, Tae; Rice, Luke M; Hancock, William O.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 15, 09.04.2019, p. 7314-7322.

Research output: Contribution to journalArticle

Mickolajczyk, Keith J. ; Geyer, Elisabeth A. ; Kim, Tae ; Rice, Luke M ; Hancock, William O. / Direct observation of individual tubulin dimers binding to growing microtubules. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 15. pp. 7314-7322.
@article{b47a092d48f64ac89a7a6316f5107ab6,
title = "Direct observation of individual tubulin dimers binding to growing microtubules",
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.",
keywords = "Interferometric scattering microscopy, Microtubule dynamics, Single-molecule kinetics",
author = "Mickolajczyk, {Keith J.} and Geyer, {Elisabeth A.} and Tae Kim and Rice, {Luke M} and Hancock, {William O.}",
year = "2019",
month = "4",
day = "9",
doi = "10.1073/pnas.1815823116",
language = "English (US)",
volume = "116",
pages = "7314--7322",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - Direct observation of individual tubulin dimers binding to growing microtubules

AU - Mickolajczyk, Keith J.

AU - Geyer, Elisabeth A.

AU - Kim, Tae

AU - Rice, Luke M

AU - Hancock, William O.

PY - 2019/4/9

Y1 - 2019/4/9

N2 - 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.

AB - 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.

KW - Interferometric scattering microscopy

KW - Microtubule dynamics

KW - Single-molecule kinetics

UR - http://www.scopus.com/inward/record.url?scp=85064110071&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85064110071&partnerID=8YFLogxK

U2 - 10.1073/pnas.1815823116

DO - 10.1073/pnas.1815823116

M3 - Article

VL - 116

SP - 7314

EP - 7322

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -