In vivo analysis of mtDNA replication defects in yeast

The yeast Saccharomyces cerevisiae has the capacity to survive large deletions or total loss of mtDNA ( petite mutants), and thus in the last few years it has been used as a model system to study defects in mitochondrial DNA (mtDNA) maintenance produced by mutations in genes involved in mtDNA replic...

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Bibliographic Details
Published in:Methods (San Diego, Calif.) Calif.), 2010-08, Vol.51 (4), p.426-436
Main Authors: Baruffini, Enrico, Ferrero, Iliana, Foury, Françoise
Format: Article
Language:English
Subjects:
Base Sequence
DNA Primers - genetics
DNA Replication - genetics
DNA, Fungal - biosynthesis
DNA, Fungal - genetics
DNA, Mitochondrial - biosynthesis
DNA, Mitochondrial - genetics
Fluorescent Dyes
Genes, Fungal
Genetic Techniques
Indoles
mtDNA point mutability
mtDNA replication
Mutagenesis
Mutation
Petite frequency
rho −/rho 0
Saccharomyces cerevisiae
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Staining and Labeling
Yeast Saccharomyces cerevisiae
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Summary:The yeast Saccharomyces cerevisiae has the capacity to survive large deletions or total loss of mtDNA ( petite mutants), and thus in the last few years it has been used as a model system to study defects in mitochondrial DNA (mtDNA) maintenance produced by mutations in genes involved in mtDNA replication. In this paper we describe methods to obtain strains harboring mutations in nuclear genes essential for the integrity of mtDNA, to measure the frequency and the nature of petite mutants, to estimate the point mutation frequency in mtDNA and to determine whether a nuclear mutation is recessive or dominant and, in the latter case, the kind of dominance.
ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2010.02.023