Cell wall synthesis and central carbohydrate metabolism are interconnected by the SNF1/Mig1 pathway in Kluyveromyces lactis

Autor(en): Rippert, Dorthe 
Backhaus, Katja
Rodicio, Rosaura
Heinisch, Juergen J.
Stichwörter: BUDDING YEAST; Carbohydrate signaling; Cell Biology; Cell wall integrity; GENE-EXPRESSION; Glycolysis; INTEGRITY SENSORS; MIG1 GLUCOSE REPRESSOR; Milk yeast; MOLECULAR-GENETICS; MUTANTS; NUCLEOCYTOPLASMIC DISTRIBUTION; PLASMA-MEMBRANE MICRODOMAINS; SNF1 PROTEIN-KINASE; YEAST SACCHAROMYCES-CEREVISIAE
Erscheinungsdatum: 2017
Herausgeber: ELSEVIER GMBH, URBAN & FISCHER VERLAG
Journal: EUROPEAN JOURNAL OF CELL BIOLOGY
Volumen: 96
Ausgabe: 1
Startseite: 70
Seitenende: 81
Zusammenfassung: 
The trimeric AMP-activated kinase complex (AMPK) is conserved from yeast to humans and is best known for its role in balancing energy metabolism. Additional functions, including the regulation of cell wall biosynthesis, have been proposed for the SNF1 complex, the baker's yeast homolog of AMPK. We here demonstrate that this function is conserved in the Crabtree-negative milk yeast Kluyveromyces lactis. Deletion mutants in the genes encoding the subunits of the trimeric complex (Klsnf1, K1ga183, lasnf4) displayed increased sensitivities towards cell wall stress agents and a mutant lacking the kinase subunit had a thinner cell wall in transmission electron micrographs as compared to wild type. Epistasis analyses demonstrated that the KlSNIF1 complex acts in parallel to cell wall integrity (CWI) signaling and stress sensitivities of Klsnfl deletions can be suppressed by additional deletions in glycolytic genes (KlPFIC1, KlPFK2, KlPGI1) or by a Klmig1 mutant. Western blot analyses of an HA-tagged KlMig1 p revealed its phosphorylation on ethanol medium similar to its S. cerevisiae ortholog, but a substantial amount of protein remained phosphorylated even with high glucose concentrations. Application of cell wall stress shifted this equilibrium towards the non-phosphorylated fraction of KlMig1p. We conclude that KlMig1 p may exert a negative regulatory function on cell wall biosynthesis. (C) 2016 Elsevier GmbH. All rights reserved.
ISSN: 01719335
DOI: 10.1016/j.ejcb.2016.12.004

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