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F 1 ‐catalysed ATP hydrolysis is required for mitochondrial biogenesis in Saccharomyces cerevisiae growing under conditions where it cannot respire
Author(s) -
LefebvreLegendre Linnka,
Balguerie Axelle,
DuvezinCaubet Stéphane,
Giraud MarieFrance,
Slonimski Piotr P.,
Di Rago JeanPaul
Publication year - 2003
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.2003.03371.x
Subject(s) - biology , saccharomyces cerevisiae , biochemistry , antimycin a , atp hydrolysis , mitochondrion , ethidium bromide , electrochemical gradient , chemiosmosis , atp synthase , adenine nucleotide , atpase , yeast , nucleotide , microbiology and biotechnology , enzyme , dna , gene , membrane
Summary Mutant strains of yeast Saccharomyces cerevisiae lacking a functional F 1 ‐ATPase were found to grow very poorly under anaerobic conditions. A single amino acid replacement (K222 > E222) that locally disrupts the adenine nucleotide catalytic site in the β‐ F 1 subunit was sufficient to compromise anaerobic growth. This mutation also affected growth in aerated conditions when ethidium bromide (an intercalating agent impairing mtDNA propagation) or antimycin (an inhibitor of respiration) was included in the medium. F 1 ‐deficient cells forced to grow in oxygen‐limited conditions were shown to lose their mtDNA completely and to accumulate Hsp60p mainly under its precursor form. Fluorescence microscopy analyses with a modified GFP containing a mitochondrial targeting presequence revealed that aerobically growing F 1 ‐deficient cells stopped importing the GFP when antimycin was added to the medium. Finally, after total inactivation of the catalytic α 3 β 3 subcomplex of F 1 , mitochondria could no longer be energized by externally added ATP because of either a block in assembly or local disruption of the adenine nucleotide processing site. Altogether these data strengthen the notion that in the absence of respiration, and whether the proton translocating domain (F 0 ) of complex V is present or not, F 1 ‐catalysed hydrolysis of ATP is essential for the occurrence of vital cellular processes depending on the maintenance of an electrochemical potential across the mitochondrial inner membrane.