Properties of yeastcells depleted of the OSCP subunit of mitochondrial ATP synthase by regulated expression of the ATP5 gene

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Abstract

OSCP is a subunit of the F(A) stalk sector of yeast mitochondrial ATP synthase complex. Cells of a null mutant for OSCP, constructed by disruption of the chromosomal ATP5 gene of Saccharomyces cerevisiae, exhibited a high level of genetic instability (petite formation). Study of the effects of ablation of OSCP required the development of a progressive depletion strategy. Introduction of a vector bearing an ATP5 gene cassette under GAL1 transcriptional control into null mutant cells gave rise to a stable yeast strain from which OSCP could be depleted in a controlled manner by manipulation of the level of galactose in the growth medium. Cells progressively depleted of OSCP exhibited properties of cellular respiration indicative of a decline in the functional coupling of the catalytic F1 sector to the proton channel F0 sector (normally linked by F(A)). Cells depleted of OSCP also exhibited a physical uncoupling of F1 from other subunits of the complex such that other F(A) subunits and F0 subunit 6 were not recovered in immunoprecipitates of ATP synthase complexes. Thus, OSCP plays a role in the assembly as well as function of the enzyme complex.

Original languageEnglish
Pages (from-to)789-799
Number of pages11
JournalBiochemistry and Molecular Biology International
Volume34
Issue number4
Publication statusPublished - 1 Jan 1994

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