Rescue of Yeast Defective in Mitochondrial ATP Synthase Subunit 8 by a Heterologous Gene From Aspergillus Nidulans

Biochem Biophys Res Commun. 1994 Sep 30;203(3):1567-73. doi: 10.1006/bbrc.1994.2365.

Abstract

Mitochondrial ATP synthase subunit 8 of the yeast Saccharomyces cerevisiae and of the filamentous fungus Aspergillus nidulans have the same length and similar structural motifs. However, the two proteins share only 50% identical residues, with the conserved residues being concentrated in the N- and C-terminal domains. We have investigated whether it is amino acid sequence or overall structural motifs that are required for subunit 8 function. PCR was used to construct a gene encoding A. nidulans subunit 8 fused to an N-terminal cleavable mitochondrial targeting sequence. Following expression in the nucleus of a yeast strain deficient in subunit 8, the chimaeric precursor targeted the subunit 8 protein back to the mitochondrion. The A. nidulans subunit 8 was found to be able to restore growth on non-fermentable substrate at 18 degrees C and 28 degrees C, but not at 36 degrees C. Given the sequence divergence between subunit 8 of A. nidulans and that of S. cerevisiae, this finding suggests that common structural motifs are important for subunit 8 function.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus nidulans / enzymology*
  • Aspergillus nidulans / genetics
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • Escherichia coli
  • Gene Deletion*
  • Genes, Fungal*
  • Macromolecular Substances
  • Mitochondria / enzymology*
  • Plasmids
  • Polymerase Chain Reaction
  • Proton-Translocating ATPases / biosynthesis
  • Proton-Translocating ATPases / genetics*
  • Restriction Mapping
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • Macromolecular Substances
  • Proton-Translocating ATPases