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Expression and evolution of the non-canonically translated yeast mitochondrial acetyl-CoA carboxylase Hfa1p.

Suomi F, Menger KE, Monteuuis G, Naumann U, Kursu VA, Shvetsova A, Kastaniotis AJ - PLoS ONE (2014)

Bottom Line: Our Δhfa1 baker's yeast mutant phenotype rescue studies using the protoploid Kluyveromyces lactis ACC confirmed functionality of the cryptic upstream mitochondrial targeting signal.These results lend strong experimental support to the hypothesis that the mitochondrial and cytosolic acetyl-CoA carboxylases in S. cerevisiae have evolved from a single gene encoding both the mitochondrial and cytosolic isoforms.Leaning on a cursory survey of a group of genes of our interest, we propose that cryptic 5' upstream mitochondrial targeting sequences may be more abundant in eukaryotes than anticipated thus far.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu, University of Oulu, Oulu, Finland.

ABSTRACT
The Saccharomyces cerevisiae genome encodes two sequence related acetyl-CoA carboxylases, the cytosolic Acc1p and the mitochondrial Hfa1p, required for respiratory function. Several aspects of expression of the HFA1 gene and its evolutionary origin have remained unclear. Here, we determined the HFA1 transcription initiation sites by 5' RACE analysis. Using a novel "Stop codon scanning" approach, we mapped the location of the HFA1 translation initiation site to an upstream AUU codon at position -372 relative to the annotated start codon. This upstream initiation leads to production of a mitochondrial targeting sequence preceding the ACC domains of the protein. In silico analyses of fungal ACC genes revealed conserved "cryptic" upstream mitochondrial targeting sequences in yeast species that have not undergone a whole genome duplication. Our Δhfa1 baker's yeast mutant phenotype rescue studies using the protoploid Kluyveromyces lactis ACC confirmed functionality of the cryptic upstream mitochondrial targeting signal. These results lend strong experimental support to the hypothesis that the mitochondrial and cytosolic acetyl-CoA carboxylases in S. cerevisiae have evolved from a single gene encoding both the mitochondrial and cytosolic isoforms. Leaning on a cursory survey of a group of genes of our interest, we propose that cryptic 5' upstream mitochondrial targeting sequences may be more abundant in eukaryotes than anticipated thus far.

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Acetyl-CoA carboxylases from yeast split into two subgroups with the exception of the S. cerevisiae HFA1.The phylogenetic tree of representative members of the acetyl-CoA carboxylases from fungi species was constructed using Clustal W2 Phylogeny and NJPlot as described in Material and methods. Numbers at branch points are bootstrap values and were calculated with Clustal X2.
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pone-0114738-g005: Acetyl-CoA carboxylases from yeast split into two subgroups with the exception of the S. cerevisiae HFA1.The phylogenetic tree of representative members of the acetyl-CoA carboxylases from fungi species was constructed using Clustal W2 Phylogeny and NJPlot as described in Material and methods. Numbers at branch points are bootstrap values and were calculated with Clustal X2.

Mentions: Querying databases on the amino acid sequences of fungal ACCs, we found that the MTS is highly conserved among fungal species especially in the group of Saccharomycotina (Fig. 5). The alignment shows the homologous sequence of ACCs through all the fungal species, except for the N-terminal part. Our results are presented in a distance tree, displaying in the division of the fungal ACCs in two large groups (Fig. 5). One group (group 1) includes Candida albicans, Candida dubliniensis, Candida tropicalis, and Debaryomyces hansenii, and is restricted to organisms which translate CTG as serine instead of leucine. Group 2 contains the other Saccharomyces species, Ashbya gossypii, Kluyveromyces waltii, K. lactis, and two post-whole genomic duplication species, Candida glabrata and S. cerevisiae. This grouping matches with the fungal phylogeny data reported by Fitzpatrick et al.[23][24]. S. cerevisiae Hfa1p is distantly located from the other ACCs from fungi, indicating that this protein has undergone a faster evolution compared to the other species.


Expression and evolution of the non-canonically translated yeast mitochondrial acetyl-CoA carboxylase Hfa1p.

Suomi F, Menger KE, Monteuuis G, Naumann U, Kursu VA, Shvetsova A, Kastaniotis AJ - PLoS ONE (2014)

Acetyl-CoA carboxylases from yeast split into two subgroups with the exception of the S. cerevisiae HFA1.The phylogenetic tree of representative members of the acetyl-CoA carboxylases from fungi species was constructed using Clustal W2 Phylogeny and NJPlot as described in Material and methods. Numbers at branch points are bootstrap values and were calculated with Clustal X2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263661&req=5

pone-0114738-g005: Acetyl-CoA carboxylases from yeast split into two subgroups with the exception of the S. cerevisiae HFA1.The phylogenetic tree of representative members of the acetyl-CoA carboxylases from fungi species was constructed using Clustal W2 Phylogeny and NJPlot as described in Material and methods. Numbers at branch points are bootstrap values and were calculated with Clustal X2.
Mentions: Querying databases on the amino acid sequences of fungal ACCs, we found that the MTS is highly conserved among fungal species especially in the group of Saccharomycotina (Fig. 5). The alignment shows the homologous sequence of ACCs through all the fungal species, except for the N-terminal part. Our results are presented in a distance tree, displaying in the division of the fungal ACCs in two large groups (Fig. 5). One group (group 1) includes Candida albicans, Candida dubliniensis, Candida tropicalis, and Debaryomyces hansenii, and is restricted to organisms which translate CTG as serine instead of leucine. Group 2 contains the other Saccharomyces species, Ashbya gossypii, Kluyveromyces waltii, K. lactis, and two post-whole genomic duplication species, Candida glabrata and S. cerevisiae. This grouping matches with the fungal phylogeny data reported by Fitzpatrick et al.[23][24]. S. cerevisiae Hfa1p is distantly located from the other ACCs from fungi, indicating that this protein has undergone a faster evolution compared to the other species.

Bottom Line: Our Δhfa1 baker's yeast mutant phenotype rescue studies using the protoploid Kluyveromyces lactis ACC confirmed functionality of the cryptic upstream mitochondrial targeting signal.These results lend strong experimental support to the hypothesis that the mitochondrial and cytosolic acetyl-CoA carboxylases in S. cerevisiae have evolved from a single gene encoding both the mitochondrial and cytosolic isoforms.Leaning on a cursory survey of a group of genes of our interest, we propose that cryptic 5' upstream mitochondrial targeting sequences may be more abundant in eukaryotes than anticipated thus far.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu, University of Oulu, Oulu, Finland.

ABSTRACT
The Saccharomyces cerevisiae genome encodes two sequence related acetyl-CoA carboxylases, the cytosolic Acc1p and the mitochondrial Hfa1p, required for respiratory function. Several aspects of expression of the HFA1 gene and its evolutionary origin have remained unclear. Here, we determined the HFA1 transcription initiation sites by 5' RACE analysis. Using a novel "Stop codon scanning" approach, we mapped the location of the HFA1 translation initiation site to an upstream AUU codon at position -372 relative to the annotated start codon. This upstream initiation leads to production of a mitochondrial targeting sequence preceding the ACC domains of the protein. In silico analyses of fungal ACC genes revealed conserved "cryptic" upstream mitochondrial targeting sequences in yeast species that have not undergone a whole genome duplication. Our Δhfa1 baker's yeast mutant phenotype rescue studies using the protoploid Kluyveromyces lactis ACC confirmed functionality of the cryptic upstream mitochondrial targeting signal. These results lend strong experimental support to the hypothesis that the mitochondrial and cytosolic acetyl-CoA carboxylases in S. cerevisiae have evolved from a single gene encoding both the mitochondrial and cytosolic isoforms. Leaning on a cursory survey of a group of genes of our interest, we propose that cryptic 5' upstream mitochondrial targeting sequences may be more abundant in eukaryotes than anticipated thus far.

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