32 research outputs found
Polyamine Sharing between Tubulin Dimers Favours Microtubule Nucleation and Elongation via Facilitated Diffusion
We suggest for the first time that the action of multivalent cations on
microtubule dynamics can result from facilitated diffusion of GTP-tubulin to the
microtubule ends. Facilitated diffusion can promote microtubule assembly,
because, upon encountering a growing nucleus or the microtubule wall, random
GTP-tubulin sliding on their surfaces will increase the probability of
association to the target sites (nucleation sites or MT ends).
This is an original explanation for understanding the apparent discrepancy
between the high rate of microtubule elongation and the low rate of tubulin
association at the microtubule ends in the viscous cytoplasm. The mechanism of
facilitated diffusion requires an attraction force between two tubulins, which
can result from the sharing of multivalent counterions. Natural polyamines
(putrescine, spermidine, and spermine) are present in all
living cells and are potent agents to trigger tubulin self-attraction. By using
an analytical model, we analyze the implication of facilitated diffusion
mediated by polyamines on nucleation and elongation of microtubules. In
vitro experiments using pure tubulin indicate that the promotion of
microtubule assembly by polyamines is typical of facilitated diffusion. The
results presented here show that polyamines can be of particular importance for
the regulation of the microtubule network in vivo and provide
the basis for further investigations into the effects of facilitated diffusion
on cytoskeleton dynamics
Multiple uses of fibrin sealant for nervous system treatment following injury and disease
Mto2 multisite phosphorylation inactivates non-spindle microtubule nucleation complexes during mitosis
Microtubule nucleation is highly regulated during the eukaryotic cell cycle, but the underlying molecular mechanisms are largely unknown. During mitosis in fission yeast Schizosaccharomyces pombe, cytoplasmic microtubule nucleation ceases simultaneously with intranuclear mitotic spindle assembly. Cytoplasmic nucleation depends on the Mto1/2 complex, which binds and activates the γ-tubulin complex and also recruits the γ-tubulin complex to both centrosomal (spindle pole body) and non-centrosomal sites. Here we show that the Mto1/2 complex disassembles during mitosis, coincident with hyperphosphorylation of Mto2 protein. By mapping and mutating multiple Mto2 phosphorylation sites, we generate mto2-phosphomutant strains with enhanced Mto1/2 complex stability, interaction with the γ-tubulin complex and microtubule nucleation activity. A mutant with 24 phosphorylation sites mutated to alanine, mto2[24A], retains interphase-like behaviour even in mitotic cells. This provides a molecular-level understanding of how phosphorylation ‘switches off' microtubule nucleation complexes during the cell cycle and, more broadly, illuminates mechanisms regulating non-centrosomal microtubule nucleation