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Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease

The most common cause of human congenital disorders of glycosylation (CDG) are mutations in the phosphomannomutase gene PMM2, which affect protein N-linked glycosylation. The yeast gene SEC53 encodes a homolog of human PMM2. We evolved 384 populations of yeast harboring one of two human-disease-asso...

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Autores principales: Vignogna, Ryan C, Allocca, Mariateresa, Monticelli, Maria, Norris, Joy W, Steet, Richard, Perlstein, Ethan O, Andreotti, Giuseppina, Lang, Gregory I
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9578706/
https://www.ncbi.nlm.nih.gov/pubmed/36214454
http://dx.doi.org/10.7554/eLife.79346
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author Vignogna, Ryan C
Allocca, Mariateresa
Monticelli, Maria
Norris, Joy W
Steet, Richard
Perlstein, Ethan O
Andreotti, Giuseppina
Lang, Gregory I
author_facet Vignogna, Ryan C
Allocca, Mariateresa
Monticelli, Maria
Norris, Joy W
Steet, Richard
Perlstein, Ethan O
Andreotti, Giuseppina
Lang, Gregory I
author_sort Vignogna, Ryan C
collection PubMed
description The most common cause of human congenital disorders of glycosylation (CDG) are mutations in the phosphomannomutase gene PMM2, which affect protein N-linked glycosylation. The yeast gene SEC53 encodes a homolog of human PMM2. We evolved 384 populations of yeast harboring one of two human-disease-associated alleles, sec53-V238M and sec53-F126L, or wild-type SEC53. We find that after 1000 generations, most populations compensate for the slow-growth phenotype associated with the sec53 human-disease-associated alleles. Through whole-genome sequencing we identify compensatory mutations, including known SEC53 genetic interactors. We observe an enrichment of compensatory mutations in other genes whose human homologs are associated with Type 1 CDG, including PGM1, which encodes the minor isoform of phosphoglucomutase in yeast. By genetic reconstruction, we show that evolved pgm1 mutations are dominant and allele-specific genetic interactors that restore both protein glycosylation and growth of yeast harboring the sec53-V238M allele. Finally, we characterize the enzymatic activity of purified Pgm1 mutant proteins. We find that reduction, but not elimination, of Pgm1 activity best compensates for the deleterious phenotypes associated with the sec53-V238M allele. Broadly, our results demonstrate the power of experimental evolution as a tool for identifying genes and pathways that compensate for human-disease-associated alleles.
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spelling pubmed-95787062022-10-19 Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease Vignogna, Ryan C Allocca, Mariateresa Monticelli, Maria Norris, Joy W Steet, Richard Perlstein, Ethan O Andreotti, Giuseppina Lang, Gregory I eLife Evolutionary Biology The most common cause of human congenital disorders of glycosylation (CDG) are mutations in the phosphomannomutase gene PMM2, which affect protein N-linked glycosylation. The yeast gene SEC53 encodes a homolog of human PMM2. We evolved 384 populations of yeast harboring one of two human-disease-associated alleles, sec53-V238M and sec53-F126L, or wild-type SEC53. We find that after 1000 generations, most populations compensate for the slow-growth phenotype associated with the sec53 human-disease-associated alleles. Through whole-genome sequencing we identify compensatory mutations, including known SEC53 genetic interactors. We observe an enrichment of compensatory mutations in other genes whose human homologs are associated with Type 1 CDG, including PGM1, which encodes the minor isoform of phosphoglucomutase in yeast. By genetic reconstruction, we show that evolved pgm1 mutations are dominant and allele-specific genetic interactors that restore both protein glycosylation and growth of yeast harboring the sec53-V238M allele. Finally, we characterize the enzymatic activity of purified Pgm1 mutant proteins. We find that reduction, but not elimination, of Pgm1 activity best compensates for the deleterious phenotypes associated with the sec53-V238M allele. Broadly, our results demonstrate the power of experimental evolution as a tool for identifying genes and pathways that compensate for human-disease-associated alleles. eLife Sciences Publications, Ltd 2022-10-10 /pmc/articles/PMC9578706/ /pubmed/36214454 http://dx.doi.org/10.7554/eLife.79346 Text en © 2022, Vignogna et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Evolutionary Biology
Vignogna, Ryan C
Allocca, Mariateresa
Monticelli, Maria
Norris, Joy W
Steet, Richard
Perlstein, Ethan O
Andreotti, Giuseppina
Lang, Gregory I
Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title_full Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title_fullStr Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title_full_unstemmed Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title_short Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
title_sort evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease
topic Evolutionary Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9578706/
https://www.ncbi.nlm.nih.gov/pubmed/36214454
http://dx.doi.org/10.7554/eLife.79346
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