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Related Experiment Videos

Kinesin-like molecules involved in spindle formation

V I Rodionov1, V I Gelfand, G G Borisy

  • 1A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia.

Journal of Cell Science
|December 1, 1993
PubMed
Summary

Microinjection of a Drosophila kinesin heavy chain antibody disrupts mitosis in PtK1 cells, preventing bipolar spindle formation and chromosome alignment. This suggests kinesin-like molecules are crucial for mitotic spindle structure.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cytoskeleton Dynamics

Background:

  • Kinesin motor proteins play vital roles in intracellular transport and cell division.
  • The specific involvement of kinesin-like molecules in mitotic spindle organization requires further elucidation.

Purpose of the Study:

  • To investigate the role of kinesin-like molecules in mitosis using a specific antibody.
  • To determine the impact of inhibiting kinesin heavy chain function on spindle formation and chromosome dynamics.

Main Methods:

  • Microinjection of a polyclonal antibody against Drosophila kinesin heavy chain (HD antibody) into PtK1 cells.
  • Time-lapse microscopy to record cell progression through mitosis.
  • Anti-tubulin immunofluorescence and immunoblotting to analyze microtubule organization and protein interactions.

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Main Results:

  • Antibody injection prevented bipolar spindle formation, leading to a monopolar spindle configuration with chromosomes arranged in a rosette.
  • Microtubules formed a radial array around a single pole.
  • Injected cells eventually entered the next cell cycle, indicating cell cycle progression despite mitotic defects.
  • The antibody recognized a 120 kDa kinesin heavy chain and other polypeptides in cell extracts and spindle preparations.

Conclusions:

  • A kinesin-like molecule is essential for the formation and/or maintenance of the mitotic spindle structure.
  • Disruption of kinesin function leads to severe defects in chromosome congression and spindle bipolarity.
  • Kinesin-like proteins are critical regulators of mitotic progression.