The kinase Isr1 negatively regulates hexosamine biosynthesis in S. cerevisiae

Autoři: Emma B. Alme aff001;  Erica Stevenson aff003;  Nevan J. Krogan aff003;  Danielle L. Swaney aff003;  David P. Toczyski aff001
Působiště autorů: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California, United States of America aff001;  Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, California, United States of America aff002;  Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California, United States of America aff003;  California Institute for Quantitative Biosciences, University of California San Francisco, San Francisco, California, United States of America aff004;  J. David Gladstone Institutes, San Francisco, California, United States of America aff005
Vyšlo v časopise: The kinase Isr1 negatively regulates hexosamine biosynthesis in S. cerevisiae. PLoS Genet 16(6): e32767. doi:10.1371/journal.pgen.1008840
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008840


The S. cerevisiae ISR1 gene encodes a putative kinase with no ascribed function. Here, we show that Isr1 acts as a negative regulator of the highly-conserved hexosamine biosynthesis pathway (HBP), which converts glucose into uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), the carbohydrate precursor to protein glycosylation, GPI-anchor formation, and chitin biosynthesis. Overexpression of ISR1 is lethal and, at lower levels, causes sensitivity to tunicamycin and resistance to calcofluor white, implying impaired protein glycosylation and reduced chitin deposition. Gfa1 is the first enzyme in the HBP and is conserved from bacteria and yeast to humans. The lethality caused by ISR1 overexpression is rescued by co-overexpression of GFA1 or exogenous glucosamine, which bypasses GFA1’s essential function. Gfa1 is phosphorylated in an Isr1-dependent fashion and mutation of Isr1-dependent sites ameliorates the lethality associated with ISR1 overexpression. Isr1 contains a phosphodegron that is phosphorylated by Pho85 and subsequently ubiquitinated by the SCF-Cdc4 complex, largely confining Isr1 protein levels to the time of bud emergence. Mutation of this phosphodegron stabilizes Isr1 and recapitulates the overexpression phenotypes. As Pho85 is a cell cycle and nutrient responsive kinase, this tight regulation of Isr1 may serve to dynamically regulate flux through the HBP and modulate how the cell’s energy resources are converted into structural carbohydrates in response to changing cellular needs.

Klíčová slova:

Biosynthesis – Cell cycle and cell division – Cell walls – Galactose – Glucose – Hyperexpression techniques – Chitin – Phosphorylation


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