Incorporation of tubulin subunits into dimers requires GTP hydrolysis.
A toroid multisubunit complex of 800-900 kDa has been implicated in assisting protein folding of at least two cytoplasmic proteins, actin and tubulin. This process is dependent on the presence of magnesium ions and ATP hydrolysis. In vitro translation of cDNAs encoding different alpha- and beta-tubulin isotypes also gives rise to the formation of complexes of about 300 kDa. These complexes have been functionally implicated in the incorporation of tubulin monomers within the tubulin heterodimer. This work shows that, in addition to ATP hydrolysis, the incorporation of newly synthesized tubulin subunits into functional heterodimers requires GTP hydrolysis in the presence of magnesium ions. A two-step process is suggested, a first ATP-dependent step in which the 900 kDa complexes are implicated in a similar way to the step taking place in actin folding, and a second GTP-dependent step in which the 300 kDa complexes are involved in the assembly of the heterodimer.
Fontalba
A
A
Departamento de Biologia Molecular, Facultad de Medicina, Universidad de Cantabria, Spain.
Paciucci
R
R
Avila
J
J
Zabala
J C
JC
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006868:Hydrolysis; D047908:Intracellular Signaling Peptides and Proteins; D008274:Magnesium / Q000378:metabolism; D051379:Mice; D008869:Microtubule-Associated Proteins; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000378:metabolism; D011487:Protein Conformation; D017510:Protein Folding; D011817:Rabbits; D051381:Rats; D014404:Tubulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases; D056427:t-Complex Genome Region
D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D014404:Tubulin; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; C116196:PPP1R11 protein, human; D044767:Ubiquitin-Protein Ligases; D008274:Magnesium