Kruszewski 2014 Abstract MiP2014
|Complementation of nuclear-encoded proteins which maintain mitochondrial DNA in S. cerevisiae with homologous proteins from other fungi species.|
Yeast mitochondrial DNA polymerase (Mip1) and RNA polymerase (Rpo41) are nuclear-encoded proteins, crucial for proper homeostasis of mitochondria and quality of mitochondrial DNA (mtDNA). Whereas organization of mitochondrial genome is different, even between related species, mitochondrial polymerases are highly evolutionary conserved in various organisms [1,2]. Complementation of Saccharomyces cerevisiae Mip1 and Rpo41 activity with proteins from other fungal species may help to understand mechanisms of mitochondrial polymerases specificity, their interactions with substrate and the general complexity of the mitochondrial-nuclear network [3,4].
We replaced activity of endogenous Mip1 and Rpo41 (separate experiments) from S. cerevisiae with their homologs from S. bayanus, S. paradoxus, Kluyveromyces lactis, Candida glabrata and Candida albicans . We analyzed efficiencies of maintaining mtDNA quality in chimeric strains and compared them to wild type. To establish how efficiently non-native polymerases could process their substrates in S. cerevisiae, chimeric strains with homologs from other fungi species were created. Phenotype analysis of obtained strains included drop assay and petite frequencies analysis. For K. lactis, C. glabrata and C. albicans we created strains with promoters and terminators from those species and other strains with native versions from S. cerevisiae.
Our results show that it is possible to complement activity of those polymerases with homologous proteins from related species, and that promoters and terminators from another yeast species are recognized by S. cerevisiae transcription factors. Therefore, we suggest that the described chimeric strains of baker’s yeast can be used as a model for studying mutual evolution of mtDNA and nuclear-encoded proteins.
Labels: MiParea: mtDNA;mt-genetics
Organism: Saccharomyces cerevisiae, Fungi
Event: A1, P-flash MiP2014
1-Inst Genetics Biotech, Univ Warsaw; 2-Inst Biochem Biophysics, Polish Acad Sc, Warsaw; Poland. - firstname.lastname@example.org
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