Abstract
The subunit analogous to the d-subunit of ATP synthase from bovine heart mitochondria was isolated from the purified yeast enzyme. Partial protein sequences were determined by direct methods. From this information, two oligonucleotide probes were constructed and used for screening a DNA genomic bank of Saccharomyces cerevisiae. The sequence of yeast subunit d was deduced from the DNA sequence of ATP7 gene. Mature yeast subunit d is 173 amino acids long. Its NH2-terminal serine is blocked by an N-acetyl group, and the protein has no processed NH2-terminal sequence other than the removal of the initiator methionine. The protein is predominantly hydrophilic. The amino acid sequence is 22% identical and 44% homologous to bovine subunit d. A null mutant was constructed. The mutant strain was unable to grow on glycerol medium. The mutant mitochondria had no detectable oligomycin-sensitive ATPase activity, and the catalytic sector F1 was loosely bound to the membranous part. The mutant mitochondria did not contain subunit d, and the mitochondrially encoded hydrophobic subunit 6 was not present.
Highlights
From the Slnstitut de Biochimie cellulaire et Neurochimie du Centre National dlae Recherche Scientifique (CNRS), Universite de Bordeaux II, 1, rue Camille Suint Sains 33077, Bordeaux and sthe Centre de Recherche de Biochimie et de Genitique Cellulaire du CNRS,118 route de Narbonne31062, Toulouse, France
Permeation gel chromatography in the presence of 8 M urea (Fig. 1)gave a fraction enriched with a subunit having a relative molecular mass of 19,000 Dain electrophoresis and another protein migrating like OSCP
The ATP synthase of S. cereuisiae contains at least 12 different subunits
Summary
From the Slnstitut de Biochimie cellulaire et Neurochimie du Centre National dlae Recherche Scientifique (CNRS), Universite de Bordeaux II, 1, rue Camille Suint Sains 33077, Bordeaux and sthe Centre de Recherche de Biochimie et de Genitique Cellulaire du CNRS,118 route de Narbonne31062, Toulouse, France. SDS-polyacrylamide slab gel electrophoresis of ATP synthaseof eucaryotes revealed another polypeptidein the migration range of OSCP This protein named subunit d was isolated first by Walker et al (1987) from the ttIodgwhtfueeMnortisihesnisirestgeaoactecaoetnhorndxzeordibysvdnh:ymeadytrtrdohesiivirxeawboelildclhAeypaaishttcTeiaihosvnPlsyeozcpstfyroiyhecmAnnoatsTtercaheytiPicawlnoa;tssonhteirstitoihc.hsFnheTeamluhcciessiFaaismttnIaaaeFgbllnwOyyrztazta-hyinettcmeeyserpse-pAsiesrtieoeosTclptuctfPorooobonrrmltsegotynphrnpnaoaeomtsdhrAeisteeediyTdsonFnoiPntsf-oaseAtahsr.hetnarTamvdusPeacehistsbnu.ayesrsnTeepttnhohreohadbrvaseeeeesbsfdeocesrerontienfabo,beettblhdsoittseovaihnibfebnuldeeibns.achrhceteietlodeaanrtbreityaodlfmdtatioirhnteoidacscnthacayodhnondlaofdiltryrtioihosapine.slaasoTlsuftphbtoehAulenyTpipprPterismopsttthyaeidrniayne-t is embedded in the membrane and is composedof hydrophobic In yeast Saccharomyces cerevisiae SDS-polyacrylamideslab subunits forming a specific proton-conducting pathway (for gel electrophoresis revealed that four subunits of the ATP review see Senior, 1988). These enzymes are not identical in subunit The protein of M , 19,000 is associated to thecomplex, whatcomposition since two of the subunits aredifferent; 6 - and c- ever the method of preparation used (Ryrie and Gallagher, 1979;Todd and Douglas, 1981; Hadikusumo et al, 1984).We. Centre National de la Recherche Scientifique, the MinistGrede la Recherche et de1’Enseignement Supirieur, and the Etablissement Public Rigional d’Aquitaine. The purpose of the present study was to isolate the yeast subunit anditsstructural gene, and todetermine the role of this polypeptide inside the complex
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