Incorporation of GDP-tubulin during elongation of microtubules in vitro.
Removal of GDP from tubulin E-site is not obligatory for the in vitro assembly of microtubule protein in 0.5 mM GMPPCP. This assembly, which is significantly enhanced by glycerol, produces microtubules of normal morphology and with normal composition of microtubule-associated proteins (MAPs). [3H]-GDP initially present at the E-site is shown to be incorporated directly into microtubules during assembly; this incorporation, maximally 60% of the assembled polymer, is dependent upon MAPs. These results are consistent with oligomeric species composed principally of GDP-tubulin plus MAPs, being incorporated directly into microtubules. The finding that stoichiometric GTP-tubulin formation is not an essential prerequisite for microtubule assembly may have important implications for the energetics of microtubule formation.
D000818:Animals; D002417:Cattle; D004164:Diphosphonates / Q000378:metabolism; D005990:Glycerol / Q000494:pharmacology; D006150:Guanine Nucleotides / Q000378:metabolism*; D006153:Guanosine Diphosphate / Q000378:metabolism*; D006157:Guanosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000494:pharmacology / Q000031:analogs & derivatives; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D007700:Kinetics; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
D004164:Diphosphonates; D006150:Guanine Nucleotides; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D006153:Guanosine Diphosphate; D006165:Guanylyl Imidodiphosphate; C004805:5'-guanylylmethylenebisphosphonate; D006157:Guanosine Monophosphate; D006160:Guanosine Triphosphate; D005990:Glycerol