Dynamics of microtubules from erythrocyte marginal bands.
Microtubules can adjust their length by the mechanism of dynamic instability, that is by switching between phases of growth and shrinkage. Thus far this phenomenon has been studied with microtubules that contain several components, that is, a mixture of tubulin isoforms, with or without a mixture of microtubule-associated proteins (MAPs), which can act as regulators of dynamic instability. Here we concentrate on the influence of the tubulin component. We have studied MAP-free microtubules from the marginal band of avian erythrocytes and compared them with mammalian brain microtubules. The erythrocyte system was selected because it represents a naturally stable aggregate of microtubules; second, the tubulin is largely homogeneous, in contrast to brain tubulin. Qualitatively, erythrocyte microtubules show similar features as brain microtubules, but they were found to be much less dynamic. The critical concentration of elongation, and the rates of association and dissociation of tubulin are all lower than with brain microtubules. Catastrophes are rare, rescues frequent, and shrinkage slow. This means that dynamic instability can be controlled by the tubulin isotype, independently of MAPs. Moreover, the extent of dynamic behavior is highly dependent on buffer conditions. In particular, dynamic instability is strongly enhanced in phosphate buffer, both for erythrocyte marginal band and brain microtubules. The lower stability in phosphate buffer argues against the hypothesis that a cap of tubulin.GDP.Pi subunits stabilizes microtubules. The difference in dynamics between tubulin isotypes and between the two ends of microtubules is preserved in the different buffer systems.
Trinczek
B
B
Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany.
Marx
A
A
Mandelkow
E M
EM
Murphy
D B
DB
Mandelkow
E
E
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D002645:Chickens; D004912:Erythrocytes / Q000378:metabolism / Q000648:ultrastructure*; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism / Q000648:ultrastructure; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D008954:Models, Biological; D010710:Phosphates / Q000494:pharmacology; D012542:Scattering, Radiation; D013045:Species Specificity; D013552:Swine; D013816:Thermodynamics; D014404:Tubulin / Q000378:metabolism / Q000648:ultrastructure
D008869:Microtubule-Associated Proteins; D010710:Phosphates; D014404:Tubulin