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Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease

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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.

Publisher
eLife Sciences Publications, Ltd
Date Issued
2022-10-10
Language
English
Type
Genre
Form
electronic document
Media type
Creator role
Faculty
Identifier
2050-084X
Has this item been published elsewhere?
Volume
11
Vignogna, . R. C., Allocca, . M., Monticelli, . M., Norris, . J. W., Steet, . R., Perlstein, . E. O., Andreotti, . G., & Lang, . G. I. (2022). (Vols. 11). https://doi.org/10.7554/elife.79346
Vignogna, Ryan C, Mariateresa Allocca, Maria Monticelli, Joy W Norris, Richard Steet, Ethan O Perlstein, Giuseppina Andreotti, and Gregory I Lang. 2022. https://doi.org/10.7554/elife.79346.
Vignogna, Ryan C, et al. 10 Oct. 2022, https://doi.org/10.7554/elife.79346.