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#microtubule assembly

3 public questions tagged with this topic.

What is the function of augmin in microtubule assembly?

Spindle microtubule amplification cannot rely solely on centrosomes especially in large oocytes acentriolar spindles and plant cells lacking centrioles. Augmin complex discovered as eight subunit module HAUS in humans provides branching nucleation pathway essential for spindle density. Mechanism: augmin binds along side wall of preexisting mother microtubule through its microtubule binding subunits HAUS6 and HAUS8 then recruits gamma tubulin ring complex via NEDD1 and GCP WD domain adaptor through direct interaction. Daughter microtubule nucleates at shallow angle about 35 degrees retaining same polarity as mother growing outward creating branched array increasing density without new MTOC requirement. This augments kinetochore fibers and bridging fibers maintaining robust spindle. Depletion leads to weak spindles reduced microtubule number elongated spindles chromosome mis segregation and mitotic delay. Thus augmin binds existing microtubules and recruits gamma TuRC cooperatively distinct from direct GTP binding or depolymerization promoting activities previously described for MAPs. Additional regulatory inputs including phosphorylation, small GTPases, and cargo adaptors fine tune filament assembly stability and motor activity matching cellular demands during division, migration, and mechanical stress responses efficiently.

Ref: Petry et al., Cell 2013 – Augmin binds existing microtubules and recruits gamma TuRC for branching nucleation.

In microtubule assembly, what happens when the αβ-tubulin concentration is below the critical concentration?

Polymerization equilibrium of cytoskeletal filaments obeys critical concentration concept derived from actin and tubulin thermodynamics studies of reversible polymers. Binding constant for addition at plus end determined by on rate times free dimer concentration minus off rate dissociation constant. Critical concentration Cc equals off rate over on rate threshold where net growth zero and polymer mass constant. Cellular tubulin concentration about 10 micromolar generally above Cc for plus end promoting growth but may be near Cc for minus. When alpha beta tubulin pool diluted experimentally or sequestered by stathmin effective concentration falls below Cc net loss dominates filaments shrink GTP cap lost catastrophe initiated. Conversely increase above Cc favors nucleation and elongation increasing polymer mass. Unlike covalent polymers microtubules reversible so below Cc no new growth occurs and existing polymers depolymerize to restore pool to equilibrium. In vivo local tubulin gradients regulated by expression autoregulation and sequestration guide where microtubules assemble especially during mitosis where local concentration high near chromatin favoring nucleation.

Ref: Alberts et al., Molecular Biology of the Cell, Chapter 14 – Critical concentration below Cc causes depolymerization.

What is the role of GTP hydrolysis in microtubule assembly?

Microtubules do not require GTP hydrolysis to polymerize hydrolysis follows incorporation and modulates stability rather than providing assembly energy which is entropically driven. After alpha beta dimer carrying GTP on beta adds to plus end longitudinal contacts induce GTPase activity converting beta bound GTP to GDP plus inorganic phosphate retained initially as GDP Pi lattice. GDP tubulin prefers bent conformation by about 12 degrees but constrained straight within wall creating mechanical strain stored as elastic energy. As long as terminal layers retain GTP cap maintains stabilizing influence over underlying strained lattice. When addition slows or stochastic fluctuations expose GDP layers at tip strain released protofilaments splay outward lateral bonds break rapid depolymerization ensues known as catastrophe releasing stored energy. Thus hydrolysis acts as timer converting stable polymer into unstable intermediate powering dynamic instability required for exploratory search and generating polymerization force harnessed at kinetochores and cortical force generation for spindle positioning.

Ref: Desai & Mitchison, Annu Rev Cell Dev Biol 1997 – GTP hydrolysis causing depolymerization and catastrophe timer.