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Microtubule Dynamics

Volume 777 of the series Methods in Molecular Biology pp 223-233

Date:

Photoactivatable-GFP-α-Tubulin as a Tool to Study Microtubule Plus-End Turnover in Living Human Cells

  • Catarina P. SamoraAffiliated withCentre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick Email author 
  • , Andrew D. McAinshAffiliated withCentre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick

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Abstract

The development of photactivatable (PA) variants of Green fluorescent protein (GFP) has allowed the dynamics of spatially restricted protein pools within living cells to be determined. Over the last 5 years, experiments utilizing PA-GFP fused to α-tubulin have provided important insights into the mechanisms that control microtubule dynamics in living cells. In this chapter, we describe the methodology required to generate stable cell lines expressing photoactivatable-GFP-α-tubulin and to derive quantitative measurements of tubulin turnover at microtubules plus-ends in living cells.

Key words

Microtubule α-Tubulin Photoactivatable-GFP Plus-end turnover Dynamics Kinetochore