title: The 'sequential allosteric ring' mechanism in the eukaryotic chaperonin-assisted folding of actin and tubulin.
abstract: Folding to completion of actin and tubulin in the eukaryotic cytosol requires their interaction with cytosolic chaperonin CCT [chaperonin containing tailless complex polypeptide 1 (TCP-1)]. Three-dimensional reconstructions of nucleotide-free CCT complexed to either actin or tubulin show that CCT stabilizes both cytoskeletal proteins in open and quasi-folded conformations mediated through interactions that are both subunit specific and geometry dependent. Here we find that upon ATP binding, mimicked by the non-hydrolysable analog AMP-PNP (5'-adenylyl-imido-diphosphate), to both CCT-alpha-actin and CCT- beta-tubulin complexes, the chaperonin component undergoes concerted movements of the apical domains, resulting in the cavity being closed off by the helical protrusions of the eight apical domains. However, in contrast to the GroE system, generation of this closed state does not induce the release of the substrate into the chaperonin cavity, and both cytoskeletal proteins remain bound to the chaperonin apical domains. Docking of the AMP-PNP-CCT-bound conformations of alpha-actin and beta-tubulin to their respective native atomic structures suggests that both proteins have progressed towards their native states.
authors.affiliation: Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Llorca
authors.forename: J
authors.initials: J
authors.lastname: Martín-Benito
authors.forename: J
authors.initials: J
authors.lastname: Grantham
authors.forename: M
authors.initials: M
authors.lastname: Ritco-Vonsovici
authors.forename: K R
authors.initials: KR
authors.lastname: Willison
authors.forename: J L
authors.initials: JL
authors.lastname: Carrascosa
authors.forename: J M
authors.initials: JM
authors.lastname: Valpuesta
mesh_terms: D016475:3T3 Cells; D000199:Actins / Q000737:chemistry*; D000266:Adenylyl Imidodiphosphate / Q000737:chemistry; D000495:Allosteric Site; D000818:Animals; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins / Q000737:chemistry*; D005057:Eukaryotic Cells; D006801:Humans; D021621:Imaging, Three-Dimensional; D051379:Mice; D016253:Microscopy, Immunoelectron / Q000379:methods; D008958:Models, Molecular; D011487:Protein Conformation; D017510:Protein Folding; D051381:Rats; D014404:Tubulin / Q000737:chemistry*
chemical_list: D000199:Actins; C533298:CCT2 protein, human; C533315:Tcp1 protein, mouse; D014404:Tubulin; D000266:Adenylyl Imidodiphosphate; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins