title: Site directed mutagenesis of subunit 8 of yeast mitochondrial ATP synthase. Functional and import properties of a series of C-terminally truncated forms.
abstract: The function of the positively charged C-terminal region of mitochondrially encoded subunit 8 of yeast mitochondrial ATP synthase was investigated using derivatives truncated at each of the 3 positively charged residues (Arg37, Arg42 and Lys47). Each construct, allotopically expressed in the nucleus, was tested for its ability to import and assemble functionally into ATP synthase in yeast cells unable to synthesize mitochondrial subunit 8. The efficiency of import of each construct into isolated wild-type yeast mitochondria was also determined. One construct truncated at the penultimate residue of subunit 8 (Lys47) functions in vivo and shows efficient import in vitro. Thus subunit 8 can function with only two positively charged residues. The remainder of the subunit 8 variants failed to rescue in vivo. Since they all show greatly reduced or undetectable import in vitro, presumably because of the increased hydrophobic character of the subunit 8 moiety in the chimaeric precursors, the status of these variants as regards assembly and function is not clear.
authors.affiliation: Department of Biochemistry, Monash University, Clayton, Vict., Australia.
authors.forename: D
authors.initials: D
authors.lastname: Nero
authors.forename: S M
authors.initials: SM
authors.lastname: Ekkel
authors.forename: L F
authors.initials: LF
authors.lastname: Wang
authors.forename: D G
authors.initials: DG
authors.lastname: Grasso
authors.forename: P
authors.initials: P
authors.lastname: Nagley
mesh_terms: D001483:Base Sequence; D001692:Biological Transport; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D006180:Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013329:Structure-Activity Relationship
chemical_list: D006180:Proton-Translocating ATPases