Yeast cytochrome b2 gene: isolation with antibody probes

One nuclear gene controls the removal of transient pre‐sequences from two yeast proteins: one encoded by the nuclear the other by the mitochondrial genome.

“…The proteolytic processing step occurs between asparagine and aspartic acid residues (Pratje et al, 1983). A similar sequence with asparagine and glutamic acid residues (Figure 1) could be assigned for the second processing site of cytochrome b2 (Guiard and Buhler, 1984). Therefore it is interesting to analyze whether the same proteolytic activity acts on both proteins and whether the mutation pet ts2858 also affects the processing of cytochrome b2.…”

One nuclear gene controls the removal of transient pre‐sequences from two yeast proteins: one encoded by the nuclear the other by the mitochondrial genome.

“…The proteolytic processing step occurs between asparagine and aspartic acid residues (Pratje et al, 1983). A similar sequence with asparagine and glutamic acid residues (Figure 1) could be assigned for the second processing site of cytochrome b2 (Guiard and Buhler, 1984). Therefore it is interesting to analyze whether the same proteolytic activity acts on both proteins and whether the mutation pet ts2858 also affects the processing of cytochrome b2.…”

PET genes of Saccharomyces cerevisiae.

“…One already mentioned is iso-1-cytochrome c (112, 167). The second, cytochrome b2, is mitochondrial lactate dehydrogenase (66,67); mutations in the structural gene for this protein therefore prevent utilization of lactate as a substrate. Additionally, of the two yeast citrate synthases, only the mitochondrial enzyme encoded by CITJ (171) is necessary for growth on acetate (77).…”

Structure, expression and regulation of a nuclear gene encoding a mitochondrial protein: the yeast L(+)‐lactate cytochrome c oxidoreductase (cytochrome b2).

“…To isolate the complete gene encoding yeast cytochrome b2, I benefited from the characterization of a 3-kb DNA fragment carrying part of the gene (Guiard and Buhler, 1984). This fragment, inserted into pBR322 yielded the recombinant plasmid pGB2 11 (Figure lA) which was used as a probe.…”

“…Analytical procedures Proteins were separated electrophoretically on SDS-polyacrylamide gels (Laemmli, 1970) and then electrophoretically transferred to nitrocellulose sheets (Towbin et al, 1979). Immunological detection was carried out as described (Guiard and Buhler, 1984). Preparation and screening of the yeast genomic library…”

“…The library was screened by a standard plaque hybridization protocol (Maniatis et al, 1982). The recombinant plasmid pGB211 (Guiard and Buhler, 1984), was nick-translated and used as a probe. DNA preparation Phages were prepared according to Yamamoto et al (1970) and purified by two cycles of CsCI equilibrium centrifugation.…”

One nuclear gene controls the removal of transient pre‐sequences from two yeast proteins: one encoded by the nuclear the other by the mitochondrial genome.

“…The proteolytic processing step occurs between asparagine and aspartic acid residues (Pratje et al, 1983). A similar sequence with asparagine and glutamic acid residues (Figure 1) could be assigned for the second processing site of cytochrome b2 (Guiard and Buhler, 1984). Therefore it is interesting to analyze whether the same proteolytic activity acts on both proteins and whether the mutation pet ts2858 also affects the processing of cytochrome b2.…”

PET genes of Saccharomyces cerevisiae.

“…One already mentioned is iso-1-cytochrome c (112, 167). The second, cytochrome b2, is mitochondrial lactate dehydrogenase (66,67); mutations in the structural gene for this protein therefore prevent utilization of lactate as a substrate. Additionally, of the two yeast citrate synthases, only the mitochondrial enzyme encoded by CITJ (171) is necessary for growth on acetate (77).…”

Structure, expression and regulation of a nuclear gene encoding a mitochondrial protein: the yeast L(+)‐lactate cytochrome c oxidoreductase (cytochrome b2).

“…To isolate the complete gene encoding yeast cytochrome b2, I benefited from the characterization of a 3-kb DNA fragment carrying part of the gene (Guiard and Buhler, 1984). This fragment, inserted into pBR322 yielded the recombinant plasmid pGB2 11 (Figure lA) which was used as a probe.…”

“…Analytical procedures Proteins were separated electrophoretically on SDS-polyacrylamide gels (Laemmli, 1970) and then electrophoretically transferred to nitrocellulose sheets (Towbin et al, 1979). Immunological detection was carried out as described (Guiard and Buhler, 1984). Preparation and screening of the yeast genomic library…”

“…The library was screened by a standard plaque hybridization protocol (Maniatis et al, 1982). The recombinant plasmid pGB211 (Guiard and Buhler, 1984), was nick-translated and used as a probe. DNA preparation Phages were prepared according to Yamamoto et al (1970) and purified by two cycles of CsCI equilibrium centrifugation.…”

One nuclear gene controls the removal of transient pre‐sequences from two yeast proteins: one encoded by the nuclear the other by the mitochondrial genome.

“…The proteolytic processing step occurs between asparagine and aspartic acid residues (Pratje et al, 1983). A similar sequence with asparagine and glutamic acid residues (Figure 1) could be assigned for the second processing site of cytochrome b2 (Guiard and Buhler, 1984). Therefore it is interesting to analyze whether the same proteolytic activity acts on both proteins and whether the mutation pet ts2858 also affects the processing of cytochrome b2.…”

PET genes of Saccharomyces cerevisiae.

“…One already mentioned is iso-1-cytochrome c (112, 167). The second, cytochrome b2, is mitochondrial lactate dehydrogenase (66,67); mutations in the structural gene for this protein therefore prevent utilization of lactate as a substrate. Additionally, of the two yeast citrate synthases, only the mitochondrial enzyme encoded by CITJ (171) is necessary for growth on acetate (77).…”

Structure, expression and regulation of a nuclear gene encoding a mitochondrial protein: the yeast L(+)‐lactate cytochrome c oxidoreductase (cytochrome b2).

“…To isolate the complete gene encoding yeast cytochrome b2, I benefited from the characterization of a 3-kb DNA fragment carrying part of the gene (Guiard and Buhler, 1984). This fragment, inserted into pBR322 yielded the recombinant plasmid pGB2 11 (Figure lA) which was used as a probe.…”

“…Analytical procedures Proteins were separated electrophoretically on SDS-polyacrylamide gels (Laemmli, 1970) and then electrophoretically transferred to nitrocellulose sheets (Towbin et al, 1979). Immunological detection was carried out as described (Guiard and Buhler, 1984). Preparation and screening of the yeast genomic library…”

“…The library was screened by a standard plaque hybridization protocol (Maniatis et al, 1982). The recombinant plasmid pGB211 (Guiard and Buhler, 1984), was nick-translated and used as a probe. DNA preparation Phages were prepared according to Yamamoto et al (1970) and purified by two cycles of CsCI equilibrium centrifugation.…”