BIO SCI D103 Lecture Notes - Lecture 7: Microtubule Nucleation, Pericentriolar Material, Tubulin
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24 Jul 2019
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Microtubules are polymers of the protein tubulin. Tubulin: heterodimer when a-tubulin and b-tubulin tightly bound together by noncovalent bonds. Microtubules are hollow and cylindrical built from 13 parallel protofilaments each composed of ab-tubulin heterodimers stacked head to tail. Microtubule lattice has distinct structural polarity with a-tubulins exposed at the minus end and beta at the plus. If filament is growing rapidly, the tip of the polymer is is t form, forming the gtp cap. If subunit addition is low, tip remains in d form. Transformation is sudden and random when hydrolysis catches up with rate of addition and gtp form gdp form. Dynamic instability: rapid inter-conversion between a growing and shrinking state at a uniform free subunit concentration: catastrophe: growth to shrinkage, rescue: shrinkage to growth. Microtubule organizing center (mtoc): specific intracellular location microtubules are generally nucleated from. Microtubule associated proteins (maps): proteins that bind to microtubules to stabilize them. Kinesin 13: catastrophe factor at plus end, prying protofilaments apart.
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Activity: Dynamic Instability of Microtubules
Part A
What causes catastrophe of the microtubule in vitro?
the lack of tubulin heterodimers |
non-motor Microtubule Associated Proteins (MAPs) |
GTP hydrolysis |
mutation of the β-tubulin |
Part B
What is the role of GTP in microtubule polymerization?
GTP binds the alpha and beta tubulin subunits together to form the tubulin monomer. |
GTP hydrolysis provides the energy for the polymerization of the microtubule. |
GTP is a second messenger that signals the need for polymerization/de-polymerization. |
GTP stabilizes the tip of the microtubule, allowing more monomers to be added. |
Part C
What would happen in the treadmilling experiment if a non-hydrolyzable analogue of GTP were used?
The monomers would be unable to add to the plus end, and the microtubules would shrink until they disappeared. |
The microtubule would treadmill until the new tubulin, with non-hydrolyzable GTP, reached the minus end, and then it would only extend at the plus end. |
The microtubule would add monomers at both the plus and minus ends, growing in both directions. |
The non-hydrolyzable GTP would stabilize both ends, causing treadmilling to stop. |
Part D
What is the difference between the plus and minus ends of the microtubule in in vitro experiments?
The beta subunit of the tubulin is exposed on the minus end. |
Polymerization occurs at the plus end. |
The plus end has a lower critical concentration for tubulin heterodimers. |
Catastrophe occurs at the minus end. |
Part E
How would the drug taxol affect the in vitro dynamic instability and treadmilling experiments?
Taxol would block catastrophe at the plus end in the dynamic instability experiment but not depolymerization at the minus end in the treadmilling experiment. |
Taxol would block the addition of tubulin in both experiments, leading to a destruction of the microtubules. |
Taxol would stabilize the microtubules in both experiments, leading to polymerization without catastrophe. |
Taxol would block depolymerization at the minus end in the treadmilling experiment but not catastrophe at the plus end in the dynamic instability experiment. |