title: The alpha-subunit of the mitochondrial F(1) ATPase interacts directly with the assembly factor Atp12p.
abstract: The Atp12p protein of Saccharomyces cerevisiae is required for the assembly of the F(1) component of the mitochondrial F(1)F(0) ATP synthase. In this report, we show that the F(1) alpha-subunit co-precipitates and co-purifies with a tagged form of Atp12p adsorbed to affinity resins. Moreover, sedimentation analysis indicates that in the presence of the F(1) alpha-subunit, Atp12p behaves as a particle of higher mass than is observed in the absence of the alpha-subunit. Yeast two-hybrid screens confirm the direct association of Atp12p with the alpha-subunit and indicate that the binding site for the assembly factor lies in the nucleotide-binding domain of the alpha-subunit, between Asp133 and Leu322. These studies provide the basis for a model of F(1) assembly in which Atp12p is released from the alpha-subunit in exchange for a beta-subunit to form the interface that contains the non-catalytic adenine nucleotide-binding site.
authors.affiliation: Department of Surgery, Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Z G
authors.initials: ZG
authors.lastname: Wang
authors.forename: D
authors.initials: D
authors.lastname: Sheluho
authors.forename: D L
authors.initials: DL
authors.lastname: Gatti
authors.forename: S H
authors.initials: SH
authors.lastname: Ackerman
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002417:Cattle; D018833:Chaperonins; D017931:DNA Primers / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D024101:Mitochondrial Proteins; D008958:Models, Molecular; D018832:Molecular Chaperones; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D020798:Two-Hybrid System Techniques
chemical_list: C110382:ATP12 protein, S cerevisiae; D017931:DNA Primers; D005656:Fungal Proteins; D024101:Mitochondrial Proteins; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; D018833:Chaperonins; D006180:Proton-Translocating ATPases