Abstract

Structural Basis of Tubulin Recruitment and Assembly by Microtubule Polymerases with Tumor Overexpressed Gene (TOG) Domain Arrays

Highlights

  • Microtubules (MTs) are highly dynamic polarized polymers that perform critical and diverse cellular functions including formation of bipolar mitotic spindles, intracellular organization, and modulation of cell development and cell migration (Akhmanova and Steinmetz, 2008; Akhmanova and Steinmetz, 2015)

  • We studied the ab-tubulin binding capacities and stoichiometries of near native monomeric and dimeric Alp14, which both consist of TOG1-TOG2 arrays and differ by the presence of a C-terminal SK-rich region and dimerization coiled-coil domain

  • Since TOG1 and TOG2 domains bind ab-tubulins via narrow and mostly ionic binding interfaces, we studied the effect of a moderate increase in ionic strength (100–200 mM KCl) on ab-tubulin binding capacities of TOG1 and TOG2 domains in these arrays (Ayaz et al, 2014; Ayaz et al, 2012)

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Summary

Introduction

Microtubules (MTs) are highly dynamic polarized polymers that perform critical and diverse cellular functions including formation of bipolar mitotic spindles, intracellular organization, and modulation of cell development and cell migration (Akhmanova and Steinmetz, 2008; Akhmanova and Steinmetz, 2015). MTs are assembled from ab-tubulin heterodimers ( termed ab-tubulin), and their polymerization exhibits dynamic instability arising from guanosine triphosphate (GTP) hydrolysis in btubulins at MT ends. Polymerization of ab-tubulin and GTP hydrolysis are regulated by conserved proteins that bind at MT plus-ends or along MT lattices (Akhmanova and Steinmetz, 2008; Akhmanova and Steinmetz, 2011; Akhmanova and Steinmetz, 2015; AlBassam and Chang, 2011; Al-Bassam et al, 2010; Brouhard and Rice, 2014). The XMAP215/Stu2/ Alp MT polymerases are among the best-studied families of MT regulators. They localize to the extreme tips of MT plus-ends and accelerate ab-tubulin polymerization in eukaryotes (Akhmanova and Steinmetz, 2011; Akhmanova and Steinmetz, 2015; Al-Bassam and Chang, 2011; Maurer et al, 2014).

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