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Identification of CBS2 as a mitochondrial protein in Saccharomyces cerevisiae

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The nuclear genome encoded yeast protein CBS2 is required for translational activation of mitochondrial cytochrome b RNA. Genetic studies have shown that the target sequence of the CBS2 protein is the 5′ untranslated leader sequence of cytochrome b RNA. Here we report on the intracellular localization of CBS2. CBS2 protein, expressed in Escherichia coli and prepared from inclusion bodies, was used as an antigen to raise a polyclonal rabbit antiserum. Affinity-purified CBS2 antibodies detect a 45 kDa protein in mitochondrial lysates of wild-type cells, which is absent in a strain in which the CBS2 gene has been deleted. The protein is overexpressed in mitochondrial extracts of a transformant carrying the CBS2 gene on a high copy number plasmid, but undetectable in the post-mitochondrial supernatant. Intramitochondrial localization of CBS2 was verified by in vitro import of CBS2 protein that had been synthesized in a reticulocyte lysate programmed with CBS2 mRNA transcribed in vitro. Mitochondrial import of CBS2 is not accompanied by any detectable proteolytic processing.

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Correspondence to Uwe Michaelis.

Additional information

Communicated by D.M. Lonsdale

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Michaelis, U., Rödel, G. Identification of CBS2 as a mitochondrial protein in Saccharomyces cerevisiae . Molec. Gen. Genet. 223, 394–400 (1990).

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Key words

  • Yeast
  • Mitochondria
  • CBS2 antibodies
  • CBS2 protein
  • In vitro import