Probing the force generation and stepping behavior of cytoplasmic dynein

Arne Gennerich, Samara L. Reck-Peterson

Research output: Chapter in Book/Report/Conference proceedingChapter

10 Scopus citations

Abstract

Cytoplasmic dynein, which is the largest and arguably the most complex cytoskeletal motor protein, plays fundamental roles during cell division, nuclear positioning, and organelle and mRNA transport, by generating force and movement toward the minus ends of microtubules. Consequently, dynein is central to many physiological processes, and its dysfunction is implicated in human diseases. However, the molecular mechanism by which dynein produces force and movement remains poorly understood. Here, we describe the use of optical tweezers to probe the nanometer-scale motion and force generation of individual dynein molecules, and provide a hands-on protocol for how to purify cytoplasmic dynein from budding yeast in amounts sufficient for single-molecule studies.

Original languageEnglish (US)
Title of host publicationSingle Molecule Analysis
Subtitle of host publicationMethods and Protocols
EditorsErwin J.G. Peterman, Gijs J.L. Wuite
Pages63-80
Number of pages18
DOIs
StatePublished - Oct 24 2011

Publication series

NameMethods in Molecular Biology
Volume783
ISSN (Print)1064-3745

    Fingerprint

Keywords

  • Budding yeast
  • Cytoplasmic dynein
  • Microtubules
  • Molecular motors
  • Optical trapping
  • Optical tweezers
  • Saccharomyces cerevisiae
  • Single-molecule assays

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Gennerich, A., & Reck-Peterson, S. L. (2011). Probing the force generation and stepping behavior of cytoplasmic dynein. In E. J. G. Peterman, & G. J. L. Wuite (Eds.), Single Molecule Analysis: Methods and Protocols (pp. 63-80). (Methods in Molecular Biology; Vol. 783). https://doi.org/10.1007/978-1-61779-282-3_4