Ecm33 is a novel factor involved in efficient glucose uptake for nutrition‐responsive <scp>TORC</scp>1 signaling in yeast

Umekawa, Midori; Ujihara, Masato; Nakai, Daiki; Takematsu, Hiromu; Wakayama, Mamoru

doi:10.1002/1873-3468.12882

Cited by 14 publications

(12 citation statements)

References 37 publications

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“…For glucose restriction, cells were grown in YP medium containing 2 % glucose until the early stage of growth, before shifted to YP medium containing 0.05 % glucose for 5 h. At the indicated time points, cells were collected and total protein extracts were precipitated with trichloroacetic acid-acetone method as we described previously. 12) 13) The protein amount of Emi2-GFP was monitored by immunoblotting using anti-GFP antibody (sc-9996) (Santa Cruz Biotechnology, TX, USA). The endogenous tubulin was monitored as a loading control using anti-alpha Tubulin antibody (ab-184970) (Abcam, CB, UK).…”

Section: Methodsmentioning

confidence: 99%

The Emi2 Protein of <i>Saccharomyces cerevisiae</i> is a Hexokinase Expressed under Glucose Limitation

et al. 2020

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Hexokinases catalyze glucose phosphorylation at the first step in glycolysis in eukaryotes. In the budding yeast Saccharomyces cerevisiae, three enzymes for glucose phosphorylation have long been known: Hxk1, Hxk2, and Glk1. In this study, we focus on Emi2, a previously uncharacterized hexokinase-like protein of S. cerevisiae. Our data show that the recombinant Emi2 protein (rEmi2), expressed in Escherichia coli, possesses glucose-phosphorylating activity in the presence of ATP and Mg 2+. It was also found that rEmi2 phosphorylates not only glucose but also fructose, mannose and glucosamine in vitro. In addition, we examined changes in the level of endogenous Emi2 protein in S. cerevisiae in the presence or absence of glucose and a non-fermentable carbon source. We found that the expression of Emi2 protein is tightly suppressed during proliferation in high glucose, while it is strongly upregulated in response to glucose limitation and the presence of a non-fermentable carbon source. Our data suggest that the expression of the endogenous Emi2 protein in S. cerevisiae is regulated under the control of Hxk2 in response to glucose availability in the environment.

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Section: Methodsmentioning

confidence: 99%

The Emi2 Protein of <i>Saccharomyces cerevisiae</i> is a Hexokinase Expressed under Glucose Limitation

et al. 2020

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“…Previous studies have shown that TORC1, which is at least partly synchronized with PKA/Snf1, regulates autophagy as well as the Ams1 activity . We next thought to determine whether deletion of CLB4 affects TORC1 activity.…”

Section: Resultsmentioning

confidence: 99%

“…Next, we asked whether disruption of Clb4 causes dysregulation of autophagy. As well as Ams1 activity, autophagy is suppressed under nutrient‐rich conditions but induced by glucose starvation through the activity of the TORC1 and PKA/Snf1 pathways . To quantify the magnitude of autophagy, we utilized the GFP‐Atg8 processing assay, according to reported procedures .…”

Section: Resultsmentioning

confidence: 99%

“…The assay for the intracellular Ams1 activity was carried out as we established previously . Briefly, 40 OD 600 cells subjected to glucose starvation or nitrogen starvation were collected and disrupted with glass beads in 550 µL of lysis buffer [20 m m PIPES/NaOH (pH 6.8), 1 m m EDTA, 1 m m PMSF, 1% Triton X‐100].…”

Section: Methodsmentioning

confidence: 99%

“…On the other hand, PKA is activated in the presence of replete glucose and may be responsible for the inactivation of Snf1 when glucose is available . Recent reports suggest intricate yet poorly characterized crosstalk between TORC1 and PKA/Snf1 during carbon depletion . Taken together, these reports indicate that TORC1 forms a component of the cellular response to glucose availability, most likely through interplay with PKA/Snf1.…”

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Mitotic cyclin Clb4 is required for the intracellular adaptation to glucose starvation in Saccharomyces cerevisiae

et al. 2019

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Edited by Ivan SadowskiCellular homeostasis in response to glucose availability is maintained through the tight coordination of various physiological processes, including cell proliferation, transcription, and metabolism. In this study, we use the budding yeast Saccharomyces cerevisiae to identify proteins implicated in carbon source-dependent modulation of physiological processes. We find that the mitotic cyclin Clb4 is required for optimal regulation of glucose-starvation-responsive pathways through the target of rapamycin complex 1. Cells lacking Clb4 are characterized by dysregulation of autophagy and impaired modulation of cell size. Notably, cell viability after prolonged glucose starvation is severely reduced by disruption of Clb4. We conclude that Clb4, in addition to its function in the cell cycle, plays a role in the intracellular adaptation to glucose starvation.

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Aspergillus flavus GPI-anchored protein-encoding ecm33 has a role in growth, development, aflatoxin biosynthesis, and maize infection

Chang

Zhāng

Scharfenstein

et al. 2018

Appl Microbiol Biotechnol

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Many glycosylphosphatidylinositol-anchored proteins (GPI-APs) of fungi are membrane enzymes, organization components, and extracellular matrix adhesins. We analyzed eight Aspergillus flavus transcriptome sets for the GPI-AP gene family and identified AFLA_040110, AFLA_063860, and AFLA_113120 to be among the top 5 highly expressed genes of the 36 family genes analyzed. Disruption of the former two genes did not drastically affect A. flavus growth and development. In contrast, disruption of AFLA_113120, an orthologue of Saccharomyces cerevisiae ECM33, caused a significant decrease in vegetative growth and conidiation, promoted sclerotial production, and altered conidial pigmentation. The A. flavus ecm33 null mutant, compared with the wild type and the complemented strain, produced predominantly aflatoxin B but accumulated comparable amounts of cyclopiazonic acid. It showed decreased sensitivity to Congo red at low concentrations (25-50 μg/mL) but had increased sensitivity to calcofluor white at high concentrations (250-500 μg/mL). Analyses of cell wall carbohydrates indicated that the α-glucan content was decreased significantly (p < 0.05), but the contents of chitin and ß-glucan were increased in the mutant strain. In a maize colonization study, the mutant was shown to be impaired in its infectivity and produced 3- to 4-fold lower amounts of conidia than the wild type and the complemented strain. A. flavus Ecm33 is required for proper cell wall composition and plays an important role in normal fungal growth and development, aflatoxin biosynthesis, and seed colonization.

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Ecm33 is a novel factor involved in efficient glucose uptake for nutrition‐responsive TORC1 signaling in yeast

Cited by 14 publications

References 37 publications

The Emi2 Protein of <i>Saccharomyces cerevisiae</i> is a Hexokinase Expressed under Glucose Limitation

The Emi2 Protein of <i>Saccharomyces cerevisiae</i> is a Hexokinase Expressed under Glucose Limitation

Mitotic cyclin Clb4 is required for the intracellular adaptation to glucose starvation in Saccharomyces cerevisiae

Aspergillus flavus GPI-anchored protein-encoding ecm33 has a role in growth, development, aflatoxin biosynthesis, and maize infection

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