Related Experiment Video

Updated: Mar 31, 2026

Spatiotemporal Analysis of Cytokinetic Events in Fission Yeast
11:19

Spatiotemporal Analysis of Cytokinetic Events in Fission Yeast

Published on: February 20, 2017

7.8K

Measurements of Myosin-II Motor Activity During Cytokinesis in Fission Yeast

Qing Tang1, Luther W Pollard1, Matthew Lord2

  • 1Department of Molecular Physiology & Biophysics, University of Vermont, Burlington, VT, 05405, USA.

Methods in Molecular Biology (Clifton, N.J.)
|November 1, 2015
PubMed

Insights

Fission yeast myosin-II (Myo2p) is crucial for cell division. This study presents three methods to measure its motor function, combining in vivo and in vitro approaches for a comprehensive understanding.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Motors

Background:

  • Fission yeast myosin-II (Myo2p) is essential for cytokinesis.
  • Actomyosin ring assembly and constriction rely on Myo2p motor function.

Purpose of the Study:

  • To detail three distinct methods for measuring Myo2p motor function.
  • To combine in vivo and in vitro approaches for studying myosin-II during cytokinesis.

Main Methods:

  • Actin-activated ATPases assay for bulk actomyosin motor activity.
  • Actin filament gliding assays to measure myosin-driven filament speed.
  • Myosin-bead motility assays to assess myosin ensemble speed along actin.

Main Results:

  • Established three complementary methods to quantify Myo2p motor activity.
  • Provided a framework for integrating in vivo and in vitro analyses of Myo2p.
  • Enabled detailed mechanistic insights into myosin-II function.

Conclusions:

  • The presented methods offer a robust toolkit for dissecting fission yeast myosin-II function.
  • Combining biochemical assays with established in vivo techniques enhances understanding of cytokinesis.
  • Further mechanistic studies of myosin-II in cell division are facilitated.
Keywords:
Actin -activated ATPase assayActin filament motility assayColorimetric assayEpifluorescence microscopyMyosin-bead motility assay sProtein-fluorophore sTotal internal reflection fluorescence microscopy

More Related Videos

Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy12:04

Examination of Mitotic and Meiotic Fission Yeast Nuclear Dynamics by Fluorescence Live-cell Microscopy

Published on: June 24, 2019

10.9K
Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells10:46

Motility of Single Molecules and Clusters of Bi-Directional Kinesin-5 Cin8 Purified from S. cerevisiae Cells

Published on: February 2, 2022

3.1K

Related Concept Videos

Role of Myosin in Cell Migration01:18

Role of Myosin in Cell Migration

Myosins are multimeric motor proteins involved in various cellular processes such as migration, adhesion, and proliferation. Myosin II is the most common type in animal cells, which binds and cross-links actin filaments.
Myosin II  is a hexamer comprising two heavy chains with globular heads and coiled-coil tails, two regulatory light chains, and two essential light chains. The ATPase sites on the myosin heads hydrolyze ATP, and the released phosphate generates the force for contraction....
3.7K
The Contractile Ring02:15

The Contractile Ring

Contractile rings are composed of microfilaments and are responsible for separating the daughter cells during cytokinesis. Contractile ring assembly proceeds along with other cell cycle events; however, very few mechanistic details are known about the timing and coordination of the contractile rings with the cell cycle.
A small GTPase, RhoA, controls the function and assembly of the contractile ring. RhoA belongs to the Ras superfamily of proteins. The activation of formins by RhoA promotes...
7.5K
Actin Polymerization and Cell Motility01:13

Actin Polymerization and Cell Motility

Actin is a family of globular proteins that are highly abundant in eukaryotic cells. It makes up approximately 1-5% of total cell protein concentration. Actin monomers polymerize to form a complex network of polarized filaments, the actin cytoskeleton, that plays a crucial role in many cellular processes, including cell motility, division, endocytosis, and metastasis of cancer cells.
Actin cytoskeleton dynamics can produce pushing, pulling, and resistance forces that help the cell to migrate....
7.2K
Overview of Myosin Structure and Function01:15

Overview of Myosin Structure and Function

Myosins are a family of molecular motor proteins, first identified in the skeletal muscles, where they are responsible for muscle contraction. Along with their role in muscle contraction, these proteins also play a role in the intracellular transport of molecules and vesicles. There are twenty-four classes of myosins based on their domain sequence and organization. Of the twenty-four, six classes (Myosin I, Myosin II, Myosin V, Myosin VI, Myosin VII, and Myosin X)  have been well...
7.4K
Studying the Cytoskeleton01:17

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
10.5K