Microtubule dynamics and microtubule caps: a time-resolved cryo-electron microscopy study.
Microtubules display the unique property of dynamic instability characterized by phase changes between growth and shrinkage, even in constant environmental conditions. The phases can be synchronized, leading to bulk oscillations of microtubules. To study the structural basis of dynamic instability we have examined growing, shrinking, and oscillating microtubules by time-resolved cryo-EM. In particular we have addressed three questions which are currently a matter of debate: (a) What is the relationship between microtubules, tubulin subunits, and tubulin oligomers in microtubule dynamics?; (b) How do microtubules shrink? By release of subunits or via oligomers?; and (c) Is there a conformational change at microtubule ends during the transitions from growth to shrinkage and vice versa? The results show that (a) oscillating microtubules coexist with a substantial fraction of oligomers, even at a maximum of microtubule assembly; (b) microtubules disassemble primarily into oligomers; and (c) the ends of growing microtubules have straight protofilaments, shrinking microtubules have protofilaments coiled inside out. This is interpreted as a transition from a tense to a relaxed conformation which could be used to perform work, as suggested by some models of poleward chromosome movement during anaphase.
Mandelkow
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EM
Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany.
Mandelkow
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E
Milligan
R A
RA
D000818:Animals; D002465:Cell Movement; D002474:Cell-Free System; D005615:Freezing; D006150:Guanine Nucleotides / Q000502:physiology; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron / Q000379:methods; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure*; D009391:Nephelometry and Turbidimetry; D011108:Polymers; D013552:Swine; D014404:Tubulin / Q000502:physiology*
D006150:Guanine Nucleotides; D046911:Macromolecular Substances; D011108:Polymers; D014404:Tubulin; D008274:Magnesium