BIOLOGY 2B03 Lecture Notes - Lecture 7: Microtubule Nucleation, Colchicine, Immunoglobulin Light Chain
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1 Sep 2020
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Microtubule is a tube comprised of 13 protofilaments arrayed in a circular pattern to form a very strong wall. Basic subunits of microtubules are dimers of alpha and beta tubulin proteins. Microtubule subunit that is added onto filament is a "tubulin heterodimer of alpha and beta" Alpha subunit is tightly bound to gtp and is never hydrolyzed while beta subunit is less tightly bound to gtp, and is cyclically hydrolyzed to gdp. Alpha and beta subunits are added and removed as heterodimers to the filaments. Alpha-beta-gtp has a much higher affinity for filament than alpha -beta- Microtubules are polar filaments and thus have different characteristics and dynamics at each end (+) end is fast growing while (-) end is slow-growing. The dimers are oriented such that the beta-subunit is closer to the (+) end than the alpha subunits. Dimers containing alpha-beta-gtp are added to the (+) end of growing filament (rescue) while dimers containing alpha -beta-gdp are released (catastrophe)
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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 ĆĀ²-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. |