3.6.5.6	C13A	site-directed mutagenesis, mutation of Cyst13 to Ala (GFP-A13gamma-tubulinresist) impairs GTP binding to the GTPase domain, stable co-expresses of the mutated recombinant protein in gamma-tubulin sh-U2-OS cells	
3.6.5.6	C354A	site-directed mutagenesis of beta-tubulin, the mutation dramatically reduces the rate of microtubule shrinking and the frequency of catastrophe	
3.6.5.6	C354S	site-directed mutagenesis of beta-tubulin, the mutation dramatically reduces the rate of microtubule shrinking and the frequency of catastrophe	
3.6.5.6	D158A	site-directed mutagenesis, altered directional movement of FtsI and septal PG composition in FtsZmut cells	
3.6.5.6	D212G	site-directed mutagenesis, altered directional movement of FtsI and septal PG composition in FtsZmut cells	
3.6.5.6	D269A	site-directed mutagenesis	
3.6.5.6	E238A	site-directed mutagenesis	
3.6.5.6	E250A	site-directed mutagenesis, altered directional movement of FtsI and septal PG composition in FtsZmut cells	
3.6.5.6	G105S	site-directed mutagenesis	
3.6.5.6	additional information	comparison of threadmilling speed and catalytic activity of wild-type and mutant cells and FtsZ polymers, overview. FtsZ GTPase mutants change the spatial distribution pattern but not the rate of septal PG synthesis	
3.6.5.6	additional information	generation of stably or transient transfected gamma-tubulin shRNA, pEGFP-gamma-tubulinresist, pEGFP-A13-gamma-tubulinresist, gamma-tubulinsgrest, gamma-tubulinR399A-K400A-R409A sgrest, and gamma-tubulin336-451 U2OS, and MCF10A cells. Stable co-expression of gamma-tubulin sgRNA depleting the endogenous gamma-tubulin pool. Fixed U2OS cells transiently expressing gamma-tubulin sgRNA (Cas9-crispGFP) are immunofluorescence stained with an anti-MTCO2 antibody and a gamma-tubulin antibody originated in mouse. Sg-mediated knockdown of gamma-tubulin affects the activity of the mitochondria, but not the structure of the endoplasmic reticulum	
3.6.5.6	R399A/K400A/R409A	site-directed mutagenesis	
3.6.5.6	T143G	mutation in tubulin signature motif of beta-tubulin, both GTP-binding affinity and microtubule-dependent GTPase activity are reduced at least 15 fold, mutant cells have a delay in mitosis	
3.6.5.6	T238A	naturally occuring mutation, the buried mutation T238A in alphabeta-tubulin yields microtubules with dramatically reduced shrinking rate and catastrophe frequency, the mutation uncouples the tubulin conformational and GTPase cycles, revealing allosteric control of microtubule dynamics. The mutation causes these effects by suppressing a conformational change that normally occurs in response to GTP hydrolysis in the lattice, without detectably changing the conformation of unpolymerized alphabetab-tubulin. The mutation predominantly affects post-GTPase conformational and dynamic properties of microtubules. The buried T238A mutation in beta-tubulin hyperstablizes microtubules in vivo and in vitro. Mutant-induced changes in polymerization dynamics do not result from defective GTPase activity. The T238A alphabeta-tubulin undergoes spontaneous nucleation more readily than wild-type, even in the presence of a nonhydrolyzable GTP analog, GTPgammaS, phenotype, overview