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Adenomatous polyposis coli on microtubule plus ends in cell extensions can promote microtubule net growth with or

Katsuhiro Kita1, Torsten Wittmann, Inke S Näthke

  • 1Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

Molecular Biology of the Cell
|March 10, 2006
PubMed

Insights

Adenomatous polyposis coli (APC) protein stabilizes microtubules (MTs), promoting their growth. This stabilization occurs independently of EB1, suggesting distinct binding mechanisms for MT plus ends.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Cytoskeleton Dynamics

Background:

  • Adenomatous polyposis coli (APC) protein is observed on a subset of cellular microtubules (MTs).
  • This localization suggests a potential role for APC in regulating MT dynamics within cell protrusions.

Purpose of the Study:

  • To investigate the direct relationship between endogenous APC and MT dynamics in living cells.
  • To determine if APC's association with MT plus ends is dependent on its binding partner EB1.

Main Methods:

  • Simultaneous visualization of endogenous APC and MT dynamics using fluorescently labeled antibodies and tubulin in living cells.
  • Comicroinjection techniques to label proteins and track MTs.
  • Utilized APC-deficient cells and EB1 small interfering RNA (siRNA) for depletion studies.
  • Observed protein behavior on MTs stabilized with nocodazole.

Main Results:

  • MTs associated with APC exhibited prolonged growth periods and reduced catastrophe frequency compared to non-APC-decorated MTs.
  • Endogenous APC showed transient association with shortening MTs.
  • APC and EB1 localized to MT plus ends independently, with limited colocalization.
  • EB1 depletion did not affect APC's growth-stabilizing influence on MT plus ends.
  • APC remained bound to nocodazole-stabilized MTs, while EB1 did not.

Conclusions:

  • Endogenous APC association with MT plus ends directly correlates with increased MT growth stability.
  • APC's MT-stabilizing function can operate independently of its association with EB1.
  • APC and EB1 likely employ distinct mechanisms to associate with MT plus ends.

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