Full humanization of the glycolytic pathway in Saccharomyces cerevisiae

Francine J Boonekamp, Ewout Knibbe, Marcel A Vieira-Lara, Melanie Wijsman, Marijke A H Luttik, Karen van Eunen, Maxime den Ridder, Reinier Bron, Ana Maria Almonacid Suarez, Patrick van Rijn, Justina C Wolters, Martin Pabst, Jean-Marc Daran, Barbara M Bakker, Pascale Daran-Lapujade

OnderzoeksoutputAcademicpeer review

1 Citaat (Scopus)
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Samenvatting

Although transplantation of single genes in yeast plays a key role in elucidating gene functionality in metazoans, technical challenges hamper humanization of full pathways and processes. Empowered by advances in synthetic biology, this study demonstrates the feasibility and implementation of full humanization of glycolysis in yeast. Single gene and full pathway transplantation revealed the remarkable conservation of glycolytic and moonlighting functions and, combined with evolutionary strategies, brought to light context-dependent responses. Human hexokinase 1 and 2, but not 4, required mutations in their catalytic or allosteric sites for functionality in yeast, whereas hexokinase 3 was unable to complement its yeast ortholog. Comparison with human tissues cultures showed preservation of turnover numbers of human glycolytic enzymes in yeast and human cell cultures. This demonstration of transplantation of an entire essential pathway paves the way for establishment of species-, tissue-, and disease-specific metazoan models.

Originele taal-2English
Artikelnummer111010
Aantal pagina's32
TijdschriftCell reports
Volume39
Nummer van het tijdschrift13
DOI's
StatusPublished - 28-jun-2022

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