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Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
05:35

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Published on: March 3, 2016

EB1 acetylation by P300/CBP-associated factor (PCAF) ensures accurate kinetochore-microtubule interactions in mitosis

Peng Xia1, Zhikai Wang, Xing Liu

  • 1Anhui Key Laboratory of Cellular Dynamics and Hefei National Laboratory for Physical Sciences at Nanoscale, University of Science and Technology of China School of Life Sciences, Hefei 230027, China.

Proceedings of the National Academy of Sciences of the United States of America
|September 25, 2012
PubMed

Insights

P300/CBP-associated factor (PCAF) acetylates EB1, a microtubule protein, reducing microtubule stability. This dynamic acetylation is crucial for accurate kinetochore-microtubule interactions during cell division.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Microtubules are vital for cellular structure and dynamics in eukaryotes.
  • EB1 is a key protein regulating microtubule plus ends and interacting with other microtubule-associated proteins.

Purpose of the Study:

  • To investigate the role of P300/CBP-associated factor (PCAF) in microtubule regulation.
  • To elucidate the mechanism by which PCAF affects microtubule stability and dynamics.

Main Methods:

  • Utilized acetylation assays to detect EB1 modification by PCAF.
  • Employed a specific antibody for acetylated EB1 (acetyl-K220) to track its localization.
  • Observed the effects of EB1 acetylation on metaphase alignment and cell cycle progression.

Main Results:

  • PCAF acetylates EB1 at K220, destabilizing its C-terminal interaction domain.
  • EB1 K220 acetylation increases during mitosis and localizes to spindle microtubule plus ends.
  • Persistent EB1 acetylation disrupts metaphase alignment and checkpoint silencing, which can be rescued by Mad2 suppression.

Conclusions:

  • Dynamic acetylation of EB1 by PCAF is a novel mechanism regulating kinetochore-microtubule interactions in mitosis.
  • This acetylation controls the localization of microtubule plus-end tracking proteins, influencing cellular plasticity and dynamics.

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