Studies on the import into mitochondria of yeast ATP synthase subunits 8 and 9 encoded by artificial nuclear genes

Ruby H.P. Law, Leigh B. Farrell, Debra Nero, Rodney J. Devenish, Phillip Nagley

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Direct fusions have been constructed between each of subunits 8 and 9 from mitochondrial ATPase of Saccharomyces cerevisiae, proteins normally encoded inside mitochondria, and the cleavable N-terminal transit peptide from the nuclearly encoded precursor to subunit 9 of Neurospora crassa mitochondrial ATPase. The subunit 8 construct was imported efficiently into isolated yeast mitochondria and was processed at or very near the fusion point. When expressed in vivo from its artificial nuclear gene, this cytoplasmically synthesized form of subunit 8 restored the growth defects of aap1 mutants unable to produce subunit 8 inside the mitochondria. The subunit 9 construct was, however, unable to be imported into isolated mitochondria and could not, following nuclear expression in vivo, complement growth defects in mitochondrial oli1 mutants. This behaviour is contrasted with the previously demonstrated import competence of another yeast subunit 9 fusion, bearing the first five residues of mature N. crassa subunit 9 interposed between its own transit peptide and the yeast subunit 9 moiety.

Original languageEnglish
Pages (from-to)501-505
Number of pages5
JournalFEBS Letters
Volume236
Issue number2
DOIs
Publication statusPublished - 29 Aug 1988

Keywords

  • (Saccharomyces cerevisiae)
  • Gene relocation
  • Import
  • Membrane assembly
  • Mitochondrial ATPase complex
  • Site-directed mutagenesis

Cite this

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title = "Studies on the import into mitochondria of yeast ATP synthase subunits 8 and 9 encoded by artificial nuclear genes",
abstract = "Direct fusions have been constructed between each of subunits 8 and 9 from mitochondrial ATPase of Saccharomyces cerevisiae, proteins normally encoded inside mitochondria, and the cleavable N-terminal transit peptide from the nuclearly encoded precursor to subunit 9 of Neurospora crassa mitochondrial ATPase. The subunit 8 construct was imported efficiently into isolated yeast mitochondria and was processed at or very near the fusion point. When expressed in vivo from its artificial nuclear gene, this cytoplasmically synthesized form of subunit 8 restored the growth defects of aap1 mutants unable to produce subunit 8 inside the mitochondria. The subunit 9 construct was, however, unable to be imported into isolated mitochondria and could not, following nuclear expression in vivo, complement growth defects in mitochondrial oli1 mutants. This behaviour is contrasted with the previously demonstrated import competence of another yeast subunit 9 fusion, bearing the first five residues of mature N. crassa subunit 9 interposed between its own transit peptide and the yeast subunit 9 moiety.",
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Studies on the import into mitochondria of yeast ATP synthase subunits 8 and 9 encoded by artificial nuclear genes. / Law, Ruby H.P.; Farrell, Leigh B.; Nero, Debra; Devenish, Rodney J.; Nagley, Phillip.

In: FEBS Letters, Vol. 236, No. 2, 29.08.1988, p. 501-505.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Law, Ruby H.P.

AU - Farrell, Leigh B.

AU - Nero, Debra

AU - Devenish, Rodney J.

AU - Nagley, Phillip

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N2 - Direct fusions have been constructed between each of subunits 8 and 9 from mitochondrial ATPase of Saccharomyces cerevisiae, proteins normally encoded inside mitochondria, and the cleavable N-terminal transit peptide from the nuclearly encoded precursor to subunit 9 of Neurospora crassa mitochondrial ATPase. The subunit 8 construct was imported efficiently into isolated yeast mitochondria and was processed at or very near the fusion point. When expressed in vivo from its artificial nuclear gene, this cytoplasmically synthesized form of subunit 8 restored the growth defects of aap1 mutants unable to produce subunit 8 inside the mitochondria. The subunit 9 construct was, however, unable to be imported into isolated mitochondria and could not, following nuclear expression in vivo, complement growth defects in mitochondrial oli1 mutants. This behaviour is contrasted with the previously demonstrated import competence of another yeast subunit 9 fusion, bearing the first five residues of mature N. crassa subunit 9 interposed between its own transit peptide and the yeast subunit 9 moiety.

AB - Direct fusions have been constructed between each of subunits 8 and 9 from mitochondrial ATPase of Saccharomyces cerevisiae, proteins normally encoded inside mitochondria, and the cleavable N-terminal transit peptide from the nuclearly encoded precursor to subunit 9 of Neurospora crassa mitochondrial ATPase. The subunit 8 construct was imported efficiently into isolated yeast mitochondria and was processed at or very near the fusion point. When expressed in vivo from its artificial nuclear gene, this cytoplasmically synthesized form of subunit 8 restored the growth defects of aap1 mutants unable to produce subunit 8 inside the mitochondria. The subunit 9 construct was, however, unable to be imported into isolated mitochondria and could not, following nuclear expression in vivo, complement growth defects in mitochondrial oli1 mutants. This behaviour is contrasted with the previously demonstrated import competence of another yeast subunit 9 fusion, bearing the first five residues of mature N. crassa subunit 9 interposed between its own transit peptide and the yeast subunit 9 moiety.

KW - (Saccharomyces cerevisiae)

KW - Gene relocation

KW - Import

KW - Membrane assembly

KW - Mitochondrial ATPase complex

KW - Site-directed mutagenesis

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