The leader peptide of yeast Atp6p is required for efficient interaction with the Atp9p ring of the mitochondrial ATPase.
Atp6p (subunit 6) of the Saccharomyces cerevisiae mitochondrial ATPase is synthesized with an N-terminal 10-amino acid presequence that is cleaved during assembly of the complex. This study has examined the role of the Atp6p presequence in the function and assembly of the ATPase complex. Two mutants were constructed in which the codons for amino acids 2-9 or 2-10 of the Atp6p precursor were deleted from the mitochondrial ATP6 gene. The concentration of Atp6p and ATPase complex was approximately 2 times less in the mutants. The lower concentration of ATPase complex in the leaderless mutants correlated with less Atp6p complexed with the Atp9p ring of the F0 sector and with accumulation of an Atp6p-Atp8p complex that aggregated into polymers destined for eventual proteolytic elimination. We propose that the presequence either targets Atp6p to the Atp9p or signals insertion of the Atp6p precursor into a microcompartment of the membrane for more efficient interaction with the Atp9p ring. Despite the ATPase deficiency, growth of the leaderless atp6 mutants on respiratory substrates and the efficiency of oxidative phosphorylation were similar to that of wild type, indicating that the mutations did not affect the proton permeability of mitochondria.
Zeng
Xiaomei
X
Department of Biological Sciences, Columbia University, New York, New York 10027, USA.
Kucharczyk
Roza
R
di Rago
Jean-Paul
JP
Tzagoloff
Alexander
A
D002463:Cell Membrane Permeability / Q000502:physiology; D003062:Codon / Q000235:genetics; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D051336:Mitochondrial Membranes / Q000201:enzymology; D025261:Mitochondrial Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D011485:Protein Binding / Q000502:physiology; D021382:Protein Sorting Signals / Q000502:physiology*; D011522:Protons; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion
C501004:ATP6 protein, S cerevisiae; D003062:Codon; D026901:Membrane Transport Proteins; D009097:Multienzyme Complexes; D021382:Protein Sorting Signals; D011522:Protons; D029701:Saccharomyces cerevisiae Proteins; D025261:Mitochondrial Proton-Translocating ATPases; C491522:OLI1 protein, S cerevisiae