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Centromere protein F includes two sites that couple efficiently to depolymerizing microtubules.

Vladimir A Volkov1, Paula M Grissom2, Vladimir K Arzhanik3

  • 1Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Sciences, Moscow, Russia, 119991 Laboratory of Biophysics, Federal Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia, 117513 N. F. Gamaleya Research Institute for Epidemiology and Microbiology, Moscow, Russia, 123098.

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|June 24, 2015
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Centromere protein F (CENP-F) binds strongly to dynamic microtubules, aiding chromosome segregation during cell division. This protein acts as an effective coupler, generating significant force from shortening microtubules.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biophysics

Background:

  • Accurate chromosome segregation relies on stable kinetochore-microtubule attachments during mitosis.
  • Centromere protein F (CENP-F) possesses microtubule-binding domains, suggesting a role in kinetochore-microtubule interactions.

Purpose of the Study:

  • To investigate the microtubule-binding properties of CENP-F.
  • To determine CENP-F's contribution to kinetochore-microtubule attachments and force generation.

Main Methods:

  • Utilized in vitro assays to assess the binding preference of CENP-F's N-terminal domain for tubulin oligomers versus microtubule walls.
  • Examined the dynamics of CENP-F fragments on stable and dynamic microtubules.
  • Measured the force transduced by CENP-F fragments from shortening microtubules using microbead assays.

Main Results:

  • The N-terminal MT-binding domain of CENP-F showed a fivefold preference for curled tubulin oligomers over microtubule walls.
  • Both N-terminal and C-terminal CENP-F fragments bound to microtubules and tracked their shortening dynamics.
  • CENP-F fragments coupled to microbeads generated forces of 3-5 pN, with peaks exceeding 10 pN, from shortening microtubules.

Conclusions:

  • CENP-F's preferential binding to tubulin structures at microtubule plus ends likely stabilizes kinetochore attachments.
  • CENP-F functions as a potent coupler, effectively transducing force from dynamic, shortening microtubules during mitosis.