TY - JOUR
T1 - Full humanization of the glycolytic pathway in Saccharomyces cerevisiae
AU - Boonekamp, Francine J.
AU - Knibbe, Ewout
AU - Vieira-Lara, Marcel A.
AU - Wijsman, Melanie
AU - Luttik, Marijke A.H.
AU - van Eunen, Karen
AU - Ridder, Maxime den
AU - Pabst, Martin
AU - Daran, Jean Marc
AU - Daran-Lapujade, Pascale
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - adaptive laboratory evolution
KW - CP: Metabolism
KW - glycolysis
KW - hexokinase
KW - humanized yeast model
KW - moonlighting function
KW - pathway transplantation
KW - Saccharomyces cerevisiae
KW - synthetic biology
UR - http://www.scopus.com/inward/record.url?scp=85133144992&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2022.111010
DO - 10.1016/j.celrep.2022.111010
M3 - Article
C2 - 35767960
AN - SCOPUS:85133144992
SN - 2211-1247
VL - 39
SP - 111010
JO - Cell Reports
JF - Cell Reports
IS - 13
M1 - 111010
ER -