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β3 accelerates microtubule plus end maturation through a divergent lateral interface.

Lisa M Wood, Jeffrey K Moore1

  • 1Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, Aurora, CO 80045.

Molecular Biology of the Cell
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Limited sequence differences in beta-tubulin (TUBB3/β3) impact microtubule dynamics and cancer drug response. Specific lateral interface residues in β3 tubulin drive these unique activities, affecting microtubule growth and maturation.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Beta-tubulin isotypes share similar sequences but possess distinct functions.
  • The TUBB3/β3 isotype is implicated in aggressive cancers and chemotherapy resistance.
  • Functional significance of minor sequence variations between tubulin isotypes is not fully understood.

Purpose of the Study:

  • To investigate the functional importance of sequence divergence in TUBB3/β3.
  • To identify specific residues and regions in β3 tubulin responsible for its unique activity.
  • To elucidate the mechanistic basis of β3 tubulin's role in microtubule dynamics and drug resistance.

Main Methods:

  • Creation of mutant yeast strains expressing β-tubulin alleles mimicking β3 variant residues.
  • Analysis of microtubule dynamics in yeast and HeLa cells overexpressing β3.
  • Assessment of microtubule response to microtubule-targeting agents, including paclitaxel.
  • Examination of EB binding at microtubule plus ends and lattice maturation rates.

Main Results:

  • Residues at the lateral interface of β3 tubulin are sufficient to alter microtubule dynamics and response to drugs.
  • Overexpression of β3 in HeLa cells shortens microtubule growth lifetime and reduces EB binding duration.
  • β3-containing microtubules exhibit faster lattice maturation and resist paclitaxel stabilization.
  • Specific regions (H1-S2 and H2-S3) at the β3 lateral interface are critical for paclitaxel resistance.

Conclusions:

  • Sequence variations at the lateral interface of β3 tubulin mechanistically explain its distinct activity.
  • Tubulin isotype expression can modulate the rate of microtubule lattice maturation at growing plus ends.
  • Findings provide insights into the role of β3 tubulin in cancer progression and chemoresistance.