You have discovered a single cellular eukaryotic organism that has very little vesicle trafficking. You suspect that this may be due to the relatively few formed microtubules that are observed in this species. Upon close examination, you find the microtubules that are present are unusually stable. When looking at an amino acid comparison with human alpha and beta tubulin, you notice that the alpha tubulin is identical, but the beta tubulin is different in the region responsible for GTP hydrolysis. You isolated the heterodimer subunits from this new species and put them in a cell free system in a GTP bound form of the heterodimer along with centrosome nucleation sites. How would the dynamics be different from normal heterodimer subunits in the same situation?
a) The new speciesâ microtubules would treadmill at a maximum concentration of heterodimers
b)The new speciesâ microtubules would be less susceptible to catastrophe
c)The new speciesâ microtubules would loose more from the (-) end than (+) end
d)The new speciesâ microtubules would grow to a longer length than normal cellular microtubules
more than one possible answer
You have discovered a single cellular eukaryotic organism that has very little vesicle trafficking. You suspect that this may be due to the relatively few formed microtubules that are observed in this species. Upon close examination, you find the microtubules that are present are unusually stable. When looking at an amino acid comparison with human alpha and beta tubulin, you notice that the alpha tubulin is identical, but the beta tubulin is different in the region responsible for GTP hydrolysis. You isolated the heterodimer subunits from this new species and put them in a cell free system in a GTP bound form of the heterodimer along with centrosome nucleation sites. How would the dynamics be different from normal heterodimer subunits in the same situation?
a) The new speciesâ microtubules would treadmill at a maximum concentration of heterodimers
b)The new speciesâ microtubules would be less susceptible to catastrophe
c)The new speciesâ microtubules would loose more from the (-) end than (+) end
d)The new speciesâ microtubules would grow to a longer length than normal cellular microtubules
more than one possible answer
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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. |