Manipulation and quantification of microtubule lattice integrity

Taylor A. Reid, Courtney Coombes, Melissa K. Gardner

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Microtubules are structural polymers that participate in a wide range of cellular functions. The addition and loss of tubulin subunits allows the microtubule to grow and shorten, as well as to develop and repair defects and gaps in its cylindrical lattice. These lattice defects act to modulate the interactions of microtubules with molecular motors and other microtubule-associated proteins. Therefore, tools to control and measure microtubule lattice structure will be invaluable for developing a quantitative understanding of how the structural state of the microtubule lattice may regulate its interactions with other proteins. In this work, we manipulated the lattice integrity of in vitro microtubules to create pools of microtubules with common nucleotide states, but with variations in structural states. We then developed a series of novel semi-automated analysis tools for both fluorescence and electron microscopy experiments to quantify the type and severity of alterations in microtubule lattice integrity. These techniques will enable new investigations that explore the role of microtubule lattice structure in interactions with microtubule-associated proteins.

Original languageEnglish (US)
Pages (from-to)1245-1256
Number of pages12
JournalBiology Open
Volume6
Issue number8
DOIs
StatePublished - Aug 15 2017

Bibliographical note

Funding Information:
Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the NSF-funded Materials Research Facilities Network (www.mrfn.org) via the MRSEC program. The steerable line filter implementation for the automated microtubule detection code was provided by the Computational Image Analysis in Cellular and Developmental Biology course at Marine Biological Laboratories. We thank Mark McClellan for technical assistance and helpful discussions. This work was supported by the National Institutes of Health (GM-103833).

Publisher Copyright:
© 2017, Company of Biologists Ltd. All rights reserved.

Keywords

  • GMPCPP
  • Lattice
  • Microtubule
  • Structure
  • Taxol

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