The alpha-subunit of the mitochondrial F(1) ATPase interacts directly with the assembly factor Atp12p.
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.
Wang
Z G
ZG
Department of Surgery, Wayne State University School of Medicine, Detroit, MI, USA.
Sheluho
D
D
Gatti
D L
DL
Ackerman
S H
SH
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
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