Dataset Information

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient afo1 ^- Yeast Suppressed by a Dominant Mutation in ATP3.

ABSTRACT: A yeast deletion mutation in the nuclear-encoded gene, AFO1, which codes for a mitochondrial ribosomal protein, led to slow growth on glucose, the inability to grow on glycerol or ethanol, and loss of mitochondrial DNA and respiration. We noticed that afo1 ^- yeast readily obtains secondary mutations that suppress aspects of this phenotype, including its growth defect. We characterized and identified a dominant missense suppressor mutation in the ATP3 gene. Comparing isogenic slowly growing rho-zero and rapidly growing suppressed afo1 ^- strains under carefully controlled fermentation conditions showed that energy charge was not significantly different between strains and was not causal for the observed growth properties. Surprisingly, in a wild-type background, the dominant suppressor allele of ATP3 still allowed respiratory growth but increased the petite frequency. Similarly, a slow-growing respiratory deficient afo1 ^- strain displayed an about twofold increase in spontaneous frequency of point mutations (comparable to the rho-zero strain) while the suppressed strain showed mutation frequency comparable to the respiratory-competent WT strain. We conclude, that phenotypes that result from afo1 ^- are mostly explained by rapidly emerging mutations that compensate for the slow growth that typically follows respiratory deficiency.

SUBMITTER: Li J

PROVIDER: S-EPMC7718765 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient afo1- Yeast Suppressed by a Dominant Mutation in ATP3.

Li Jing J Rinnerthaler Mark M Hartl Johannes J Weber Manuela M Karl Thomas T Breitenbach-Koller Hannelore H Mülleder Michael M Vowinckel Jakob J Marx Hans H Sauer Michael M Mattanovich Diethard D Ata Özge Ö De Sonakshi S Greslehner Gregor P GP Geltinger Florian F Burhans Bill B Grant Chris C Doronina Victoria V Ralser Meryem M Streubel Maria Karolin MK Grabner Christian C Jarolim Stefanie S Moßhammer Claudia C Gourlay Campbell W CW Hasek Jiri J Cullen Paul J PJ Liti Gianni G Ralser Markus M Breitenbach Michael M

G3 (Bethesda, Md.) 20201203 12

A yeast deletion mutation in the nuclear-encoded gene, AFO1, which codes for a mitochondrial ribosomal protein, led to slow growth on glucose, the inability to grow on glycerol or ethanol, and loss of mitochondrial DNA and respiration. We noticed that afo1- yeast readily obtains secondary mutations that suppress aspects of this phenotype, including its growth defect. We characterized and identified a dominant missense suppressor mutation in the ATP3 gene. Comparin ...[more]

PMID: 33093184

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Project description:UnlabelledEbola virus (EBOV) is an RNA virus that can cause hemorrhagic fever with high fatality rates, and there are no approved vaccines or therapies. Typically, RNA viruses have high spontaneous mutation rates, which permit rapid adaptation to selection pressures and have other important biological consequences. However, it is unknown if filoviruses exhibit high mutation frequencies. Ultradeep sequencing and a recombinant EBOV that carries the gene encoding green fluorescent protein were used to determine the spontaneous mutation frequency of EBOV. The effects of the guanosine analogue ribavirin during EBOV infections were also assessed. Ultradeep sequencing revealed that the mutation frequency for EBOV was high and similar to those of other RNA viruses. Interestingly, significant genetic diversity was not observed in viable viruses, implying that changes were not well tolerated. We hypothesized that this could be exploited therapeutically. In vitro, the presence of ribavirin increased the error rate, and the 50% inhibitory concentration (IC50) was 27 μM. In a mouse model of ribavirin therapy given pre-EBOV exposure, ribavirin treatment corresponded with a significant delay in time to death and up to 75% survival. In mouse and monkey models of therapy given post-EBOV exposure, ribavirin treatment also delayed the time to death and increased survival. These results demonstrate that EBOV has a spontaneous mutation frequency similar to those of other RNA viruses. These data also suggest a potential for therapeutic use of ribavirin for human EBOV infections.ImportanceEbola virus (EBOV) causes a severe hemorrhagic disease with high case fatality rates; there are no approved vaccines or therapies. We determined the spontaneous mutation frequency of EBOV, which is relevant to understanding the potential for the virus to adapt. The frequency was similar to those of other RNA viruses. Significant genetic diversity was not observed in viable viruses, implying that changes were not well tolerated. We hypothesized that this could be exploited therapeutically. Ribavirin is a viral mutagen approved for treatment of several virus infections; it is also cheap and readily available. In cell culture, we showed that ribavirin was effective at reducing production of infectious EBOV. In mouse and monkey models of therapy given post-EBOV exposure, ribavirin treatment delayed the time to death and increased survival. These data provide a better understanding of EBOV spontaneous mutation and suggest that ribavirin may have great value in the context of human disease.

Dataset Information

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient afo1 ^- Yeast Suppressed by a Dominant Mutation in ATP3.

Publications

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient <i>afo1</i><sup>-</sup> Yeast Suppressed by a Dominant Mutation in <i>ATP3</i>.

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Dataset Information

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient afo1 - Yeast Suppressed by a Dominant Mutation in ATP3.

Publications

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient <i>afo1</i><sup>-</sup> Yeast Suppressed by a Dominant Mutation in <i>ATP3</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Slow Growth and Increased Spontaneous Mutation Frequency in Respiratory Deficient afo1 ^- Yeast Suppressed by a Dominant Mutation in ATP3.