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Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

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Published on: March 15, 2014

The human kinetochore Ska1 complex facilitates microtubule depolymerization-coupled motility.

Julie P I Welburn¹, Ekaterina L Grishchuk, Chelsea B Backer

¹Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Technology, Suite 401, Nine Cambridge Center, Cambridge, MA 02142, USA.

Developmental Cell

|March 18, 2009

Summary

This summary is machine-generated.

The human Ska1 complex, including Rama1, binds microtubules and drives chromosome segregation. This complex is crucial for kinetochore-microtubule interactions during cell division.

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Area of Science:

Cell Biology
Molecular Biology
Genetics

Background:

Mitotic chromosome segregation depends on kinetochore-microtubule interactions.
The Dam1 complex in yeast links kinetochores to microtubule dynamics, but its metazoan homolog was unknown.

Purpose of the Study:

To identify proteins involved in vertebrate kinetochore-microtubule interactions.
To characterize the function and biochemical activities of the human Ska1 complex.

Main Methods:

Isolation and characterization of the human Ska1 complex.
Depletion of Ska1 complex subunits using genetic methods.
Biochemical reconstitution and in vitro assays of Ska1 complex activity.

Main Results:

Identified a three-subunit human Ska1 complex, including Rama1, at the outer kinetochore and spindle microtubules.
Depletion of Ska1 complex subunits caused severe chromosome segregation defects.
The reconstituted Ska1 complex binds microtubules and facilitates movement along depolymerizing microtubules.

Conclusions:

The human Ska1 complex plays a critical role in chromosome segregation.
Ska1 complex interacts with dynamic microtubules at the outer kinetochore.
Rama1 subunit contributes to microtubule-stimulated oligomerization.