Manipulation and quantification of microtubule lattice integrity

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

Research output: Contribution to journalArticle

3 Citations (Scopus)

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

Fingerprint

Microtubule-Associated Proteins
Microtubules
microtubules
Crystal defects
Fluorescence microscopy
Tubulin
Crystal lattices
Electron microscopy
Polymers
Repair
Nucleotides
Defects
Proteins
Experiments
Fluorescence Microscopy
proteins
Electron Microscopy
tubulin
fluorescence microscopy
electron microscopy

Keywords

  • GMPCPP
  • Lattice
  • Microtubule
  • Structure
  • Taxol

Cite this

Manipulation and quantification of microtubule lattice integrity. / Reid, Taylor A.; Coombes, Courtney; Gardner, Melissa K.

In: Biology Open, Vol. 6, No. 8, 15.08.2017, p. 1245-1256.

Research output: Contribution to journalArticle

Reid, Taylor A. ; Coombes, Courtney ; Gardner, Melissa K. / Manipulation and quantification of microtubule lattice integrity. In: Biology Open. 2017 ; Vol. 6, No. 8. pp. 1245-1256.
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