A 14 kDa release factor is involved in GTP-dependent beta-tubulin folding.
The tubulin folding pathway is a model system to understand protein folding in the cell. It involves the interaction of several chaperones, including TCP-1 and other as yet uncharacterized factors. Release of tubulin monomers from folding intermediates (C900 and C300) and their incorporation into tubulin dimers is dependent on GTP hydrolysis, magnesium ions and release factors. In this work, we have purified to homogeneity the protein factor responsible for the release of beta-tubulin monomers from C300 complexes. It has an apparent molecular mass of 14 kDa (p14) as judged by SDS electrophoresis. The protein behaved as a dimer of about 28 kDa when analyzed by gel filtration chromatography. Furthermore, the p14-dependent release of beta-tubulin monomers from C300 complexes takes place in the presence of GTP. These results suggest that p14 is a new chaperone that assists in tubulin folding by facilitating the acquisition of the native conformation.
Campo
R
R
Departamento de Biologia Molecular, Facultad de Medicina, Universidad de Cantabria, Santander, Spain.
Fontalba
A
A
Sanchez
L M
LM
Zabala
J C
JC
D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D006868:Hydrolysis; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D008297:Male; D008869:Microtubule-Associated Proteins; D018832:Molecular Chaperones / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000378:metabolism; D017510:Protein Folding; D011817:Rabbits; D013552:Swine; D014404:Tubulin / Q000737:chemistry*; D044767:Ubiquitin-Protein Ligases; D056427:t-Complex Genome Region
D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D014404:Tubulin; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; C116196:PPP1R11 protein, human; D044767:Ubiquitin-Protein Ligases