Limits...
Post-zygotic sterility and cytonuclear compatibility limits in S. cerevisiae xenomitochondrial cybrids.

Špírek M, Poláková S, Jatzová K, Sulo P - Front Genet (2015)

Bottom Line: Their decreased respiration capacity and reduced cytochrome aa3 content is associated with the inefficient splicing of cox1I3β, the intron found in all Saccharomyces species but not in S. cerevisiae.The nucleo-mitochondrial compatibility limit of S. cerevisiae and other Saccharomyces was set between S. kudriavzevii and S. bayanus at the divergence from S. cerevisiae about 15 MYA.The MRS1-cox1 S. cerevisiae/S. paradoxus cytonuclear Dobzhansky-Muller pair has a neglible impact on the separation of species since its imperfection is compensated for by gain-of-function mutation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University Bratislava, Slovakia.

ABSTRACT
Nucleo-mitochondrial interactions, particularly those determining the primary divergence of biological species, can be studied by means of xenomitochondrial cybrids, which are cells where the original mitochondria are substituted by their counterparts from related species. Saccharomyces cerevisiae cybrids are prepared simply by the mating of the ρ(0) strain with impaired karyogamy and germinating spores from other Saccharomyces species and fall into three categories. Cybrids with compatible mitochondrial DNA (mtDNA) from Saccharomyces paradoxus CBS 432 and Saccharomyces cariocanus CBS 7994 are metabolically and genetically similar to cybrids containing mtDNA from various S. cerevisiae. Cybrids with mtDNA from other S. paradoxus strains, S. cariocanus, Saccharomyces kudriavzevii, and Saccharomyces mikatae require a period of adaptation to establish efficient oxidative phosphorylation. They exhibit a temperature-sensitive phenotype, slower growth rate on a non-fermentable carbon source and a long lag phase after the shift from glucose. Their decreased respiration capacity and reduced cytochrome aa3 content is associated with the inefficient splicing of cox1I3β, the intron found in all Saccharomyces species but not in S. cerevisiae. The splicing defect is compensated in cybrids by nuclear gain-of-function and can be alternatively suppressed by overexpression of MRP13 gene for mitochondrial ribosomal protein or the MRS2, MRS3, and MRS4 genes involved in intron splicing. S. cerevisiae with Saccharomyces bayanus mtDNA is unable to respire and the growth on ethanol-glycerol can be restored only after mating to some mit (-) strains. The nucleo-mitochondrial compatibility limit of S. cerevisiae and other Saccharomyces was set between S. kudriavzevii and S. bayanus at the divergence from S. cerevisiae about 15 MYA. The MRS1-cox1 S. cerevisiae/S. paradoxus cytonuclear Dobzhansky-Muller pair has a neglible impact on the separation of species since its imperfection is compensated for by gain-of-function mutation.

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Adaptation of cybrids. Cells from single cybrid colony were diluted, spotted on the plates with the non-fermentable carbon source (YPGE) and cultivated for 7 days at 28°C. S. cerevisiae cybrids with mtDNA from: (A)S. cerevisiae W303 1A; (B)S. cerevisiae CBS 400; (C)S. paradoxus CBS 432; (D)S. cariocanus CBS 7994; (E)S. kudriavzevii CBS 8840; (F)S. paradoxus CBS 2908; (G)S. paradoxus CBS 7400; (H)S. mikatae CBS 8839; (I)S. bayanus CBS 380; (J)S. cerevisiae MCC109 ρ0.
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Figure 3: Adaptation of cybrids. Cells from single cybrid colony were diluted, spotted on the plates with the non-fermentable carbon source (YPGE) and cultivated for 7 days at 28°C. S. cerevisiae cybrids with mtDNA from: (A)S. cerevisiae W303 1A; (B)S. cerevisiae CBS 400; (C)S. paradoxus CBS 432; (D)S. cariocanus CBS 7994; (E)S. kudriavzevii CBS 8840; (F)S. paradoxus CBS 2908; (G)S. paradoxus CBS 7400; (H)S. mikatae CBS 8839; (I)S. bayanus CBS 380; (J)S. cerevisiae MCC109 ρ0.

Mentions: Cybrids with partially re-established mitochondrial functions can be obtained on selection media containing glucose. If cells from a single cybrid colony are transferred from selection media (with glucose) on the plates with a non-fermentable carbon source such as glycerol and ethanol (YPGE), unambiguously good growth is displayed only in cybrids containing mitochondrial genome from various S. cerevisiae strains, the S. paradoxus strain CBS 432, and the S. cariocanus strain CBS 7994 (Figure 3, Table 1). Other cybrids gave rise to a mixed population that consisted mainly of poorly growing colonies. However, most of these cybrids with mtDNA from S. paradoxus CBS 7400, S. paradoxus CBS 2908, and S. mikatae CBS 8839 were capable of forming colonies on non-fermentable substrate after prolonged incubation on YPGE (Figure 3, Table 1), indicating an adaptation process. Cybrids with mt genome S. kudriavzevii CBS 8840 growing on glycerol also appeared, with a shorter but significant delay. Even after 2 months of incubation we did not observe any colony growing on glycerol among the cybrids containing mtDNA from the less-related yeasts of the S. bayanus – uvarum group.


Post-zygotic sterility and cytonuclear compatibility limits in S. cerevisiae xenomitochondrial cybrids.

Špírek M, Poláková S, Jatzová K, Sulo P - Front Genet (2015)

Adaptation of cybrids. Cells from single cybrid colony were diluted, spotted on the plates with the non-fermentable carbon source (YPGE) and cultivated for 7 days at 28°C. S. cerevisiae cybrids with mtDNA from: (A)S. cerevisiae W303 1A; (B)S. cerevisiae CBS 400; (C)S. paradoxus CBS 432; (D)S. cariocanus CBS 7994; (E)S. kudriavzevii CBS 8840; (F)S. paradoxus CBS 2908; (G)S. paradoxus CBS 7400; (H)S. mikatae CBS 8839; (I)S. bayanus CBS 380; (J)S. cerevisiae MCC109 ρ0.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4290679&req=5

Figure 3: Adaptation of cybrids. Cells from single cybrid colony were diluted, spotted on the plates with the non-fermentable carbon source (YPGE) and cultivated for 7 days at 28°C. S. cerevisiae cybrids with mtDNA from: (A)S. cerevisiae W303 1A; (B)S. cerevisiae CBS 400; (C)S. paradoxus CBS 432; (D)S. cariocanus CBS 7994; (E)S. kudriavzevii CBS 8840; (F)S. paradoxus CBS 2908; (G)S. paradoxus CBS 7400; (H)S. mikatae CBS 8839; (I)S. bayanus CBS 380; (J)S. cerevisiae MCC109 ρ0.
Mentions: Cybrids with partially re-established mitochondrial functions can be obtained on selection media containing glucose. If cells from a single cybrid colony are transferred from selection media (with glucose) on the plates with a non-fermentable carbon source such as glycerol and ethanol (YPGE), unambiguously good growth is displayed only in cybrids containing mitochondrial genome from various S. cerevisiae strains, the S. paradoxus strain CBS 432, and the S. cariocanus strain CBS 7994 (Figure 3, Table 1). Other cybrids gave rise to a mixed population that consisted mainly of poorly growing colonies. However, most of these cybrids with mtDNA from S. paradoxus CBS 7400, S. paradoxus CBS 2908, and S. mikatae CBS 8839 were capable of forming colonies on non-fermentable substrate after prolonged incubation on YPGE (Figure 3, Table 1), indicating an adaptation process. Cybrids with mt genome S. kudriavzevii CBS 8840 growing on glycerol also appeared, with a shorter but significant delay. Even after 2 months of incubation we did not observe any colony growing on glycerol among the cybrids containing mtDNA from the less-related yeasts of the S. bayanus – uvarum group.

Bottom Line: Their decreased respiration capacity and reduced cytochrome aa3 content is associated with the inefficient splicing of cox1I3β, the intron found in all Saccharomyces species but not in S. cerevisiae.The nucleo-mitochondrial compatibility limit of S. cerevisiae and other Saccharomyces was set between S. kudriavzevii and S. bayanus at the divergence from S. cerevisiae about 15 MYA.The MRS1-cox1 S. cerevisiae/S. paradoxus cytonuclear Dobzhansky-Muller pair has a neglible impact on the separation of species since its imperfection is compensated for by gain-of-function mutation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University Bratislava, Slovakia.

ABSTRACT
Nucleo-mitochondrial interactions, particularly those determining the primary divergence of biological species, can be studied by means of xenomitochondrial cybrids, which are cells where the original mitochondria are substituted by their counterparts from related species. Saccharomyces cerevisiae cybrids are prepared simply by the mating of the ρ(0) strain with impaired karyogamy and germinating spores from other Saccharomyces species and fall into three categories. Cybrids with compatible mitochondrial DNA (mtDNA) from Saccharomyces paradoxus CBS 432 and Saccharomyces cariocanus CBS 7994 are metabolically and genetically similar to cybrids containing mtDNA from various S. cerevisiae. Cybrids with mtDNA from other S. paradoxus strains, S. cariocanus, Saccharomyces kudriavzevii, and Saccharomyces mikatae require a period of adaptation to establish efficient oxidative phosphorylation. They exhibit a temperature-sensitive phenotype, slower growth rate on a non-fermentable carbon source and a long lag phase after the shift from glucose. Their decreased respiration capacity and reduced cytochrome aa3 content is associated with the inefficient splicing of cox1I3β, the intron found in all Saccharomyces species but not in S. cerevisiae. The splicing defect is compensated in cybrids by nuclear gain-of-function and can be alternatively suppressed by overexpression of MRP13 gene for mitochondrial ribosomal protein or the MRS2, MRS3, and MRS4 genes involved in intron splicing. S. cerevisiae with Saccharomyces bayanus mtDNA is unable to respire and the growth on ethanol-glycerol can be restored only after mating to some mit (-) strains. The nucleo-mitochondrial compatibility limit of S. cerevisiae and other Saccharomyces was set between S. kudriavzevii and S. bayanus at the divergence from S. cerevisiae about 15 MYA. The MRS1-cox1 S. cerevisiae/S. paradoxus cytonuclear Dobzhansky-Muller pair has a neglible impact on the separation of species since its imperfection is compensated for by gain-of-function mutation.

No MeSH data available.


Related in: MedlinePlus