Evolutionary rescue of phosphomannomutase deficiency in yeast models of human disease

Elife. 2022 Oct 10:11:e79346. doi: 10.7554/eLife.79346.

Abstract

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.

Keywords: PMM2-CDG; S. cerevisiae; congenital disorders of glycosylation; evolutionary biology; experimental evolution; genetics; genomics; humanized yeast.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Congenital Disorders of Glycosylation* / genetics
  • Congenital Disorders of Glycosylation* / metabolism
  • Humans
  • Mutant Proteins
  • Phosphoglucomutase / genetics
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins* / genetics

Substances

  • phosphomannomutase
  • Phosphoglucomutase
  • Mutant Proteins
  • SEC53 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins