Isolation and Characterization of Glucose Derepressed Invertase Mutants from<i>Schizosaccharomyces pombe</i>

Kığ, Cenk; Türkel, Sezai; Temizkan, Güler

doi:10.1271/bbb.69.2475

Cited by 20 publications

(32 citation statements)

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“…This finding is consistent with that of another study, which found reduced glucose consumption by ird mutants (Kig et al, 2005). Therefore, the elevated expression of the fbp1 gene in ird mutants (Figure 2) might be because of a lack of glucose repression.…”

Section: Ability To Escape Glucose Repression In Ird Mutantssupporting

confidence: 93%

“…pombe Lindner liquefaciens (wild type, 972h -) and glucose repression resistant constitutive invertase mutants (ird5, ird13, and ird14) (Kig et al, 2005) were used in this study. The growth medium, containing 0.5% yeast extract, 3% sucrose, and 400 µg/mL 2-deoxy-Dglucose (Rincon et al, 2001) was used for the selection of ird mutants.…”

Section: Yeast Strains and Growth Mediamentioning

confidence: 99%

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Effects of glucose sensing/signaling on oxidative stress response in glucose repression mutants of Schizosaccharomyces pombe

Palabıyık

Ghods

Uçar³

2013

Genet. Mol. Res.

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ABSTRACT. The resistant to glucose repression mutants of Schizosaccharomyces pombe (ird5, ird13, and ird14) have a high tolerance to oxidative stress induced by H 2 O 2 . In all ird mutants, the increased expression level of the fbp1 gene can be interpreted as a lack of glucose repression in these mutants. To investigate the mechanisms of the oxidative stress response in ird mutants, we analyzed the transcription of stress response-related genes, sod1, ctt1, atf1, pap1, and sty1, under stressed and non-stressed conditions. We then analyzed the phosphorylation state of the Sty1-MAP kinase in ird mutants. Our findings support the concept of an adaptive response to oxidative stress in these mutants. In addition, these results imply that either glucose signaling mechanisms leading to glucose repression and glucose utilization as an energy source are regulated apart from each other or, like Saccharomyces cerevisiae, S. pombe might have additional glucose detection systems.

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Section: Ability To Escape Glucose Repression In Ird Mutantssupporting

confidence: 93%

Section: Yeast Strains and Growth Mediamentioning

confidence: 99%

Effects of glucose sensing/signaling on oxidative stress response in glucose repression mutants of Schizosaccharomyces pombe

Palabıyık

Ghods

Uçar³

2013

Genet. Mol. Res.

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“…In Sch. pombe, however, these control mechanisms have not been analysed in detail (Tanaka et al, 1998;Hoffman, 2005;Kig et al, 2005), especially for enzymes needed for utilization of galactose as an alternative carbon source. Interestingly, Sch.…”

Section: S Suzuki and Others 710mentioning

confidence: 99%

Characterization of two different types of UDP-glucose/-galactose4-epimerase involved in galactosylation in fission yeast

et al. 2010

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Schizosaccharomyces species are currently the only known organisms with two types of genes encoding UDP-glucose/-galactose 4-epimerase, uge1 + and gal10 + . A strain deleted for uge1 + exhibited a severe galactosylation defect and a decrease in activity and in UDP-galactose content when grown in glucose-rich medium (2 % glucose), indicating that Uge1p is a major UDPglucose/-galactose 4-epimerase under these growth conditions. In contrast, gal10 + was efficiently expressed and involved in galactosylation of cell-surface proteins in low-glucose medium (0.1 % glucose and 2 % glycerol), but not in galactose-containing medium. In a uge1Dgal10D strain, the galactosylation defect was suppressed and UDP-galactose content restored to wild-type levels in galactose-containing medium. Disruption of gal7 + , encoding galactose-1-phosphate uridylyltransferase, in the uge1Dgal10D strain reversed suppression of the galactosylation defect and reduced levels of UDP-galactose, indicating that galactose is transported from the medium to the cytosol and is converted into UDP-galactose via galactose 1-phosphate by Gal7p in Sch. pombe. INTRODUCTIONGlycoproteins in the fission yeast Schizosaccharomyces pombe contain a large amount of galactose in addition to both N-and O-linked mannan (Manners & Meyer, 1977;Moreno et al., 1985;Ikeda et al., 2009;Ohashi et al., 2009), indicating that Sch. pombe is equipped with mechanisms for glycoprotein galactosylation like animal cells. Research has demonstrated that the outer chain structure of galactomannan has an a-1,2-linked galactose or pyruvylated galactose attached to a poly-a-1,6-linked mannose backbone (Gemmill & Trimble, 1998), and that the pyruvylated galactose epitope bears remarkable structural resemblance to the N-acetylneuraminic acid-linked galactose epitope found in mammalian cells (Gemmill & Trimble, 1996). These galactose residues are enzymically modified by galactosyltransferases using UDP-galactose as substrate (Andreishcheva et al., 2004; Chappell et al., 1994;Yoko-o et al., 1998). UDP-galactose is transported to the Golgi lumen from the cytosol by the Golgi-localized UDPgalactose transporter (Gms1p) (Tabuchi et al., 1997;). In Sch. pombe, the precise manner in which UDP-galactose is synthesized in the cytosol has been unclear. In the cytosol, the key enzyme for UDP-galactose synthesis is UDP-galactose/-glucose 4-epimerase, which catalyses the interconversion of UDPgalactose and UDP-D-glucose. Interestingly, Schizosaccharomyces species have two types of UDP-glucose/-galactose 4-epimerase, while other organisms have only one. It is not known why two types of epimerase are needed and how they are regulated in Sch. pombe.In Saccharomyces cerevisiae, Gal10p is the sole UDPglucose/-galactose 4-epimerase, which consists of a UDPglucose/-galactose 4-epimerase domain and a galactose mutarotase domain (Majumdar et al., 2004;Scott & Timson, 2007;Thoden & Holden, 2005), and which catalyses mutarotation of a-galactose to b-galactose. Expression of the GAL10 gene is regulated by Gal4p, a DNA...

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“…Wild type S. pombe Lindner Liquifacience (972 h -) and the glucose-repression-resistant mutant (ird11) (Kig et al, 2005) were used in this study. A selective medium consisting of 0.5% yeast extract, 3% sucrose, and 400 μg/mL 2-deoxy-D-glucose was prepared for the ird11 mutant, whereas the wild type was cultured in standard rich medium (YEL).…”

Section: Yeast Strains and Mediamentioning

confidence: 99%

“…In this study, the mechanism(s) of protection from the damaging effects of oxidative stress were investigated by using the resistant glucose repression S. pombe mutant (ird11) (Kig et al, 2005), which is affected by glucose signaling in a different manner than that caused by glucose deprivation (Palabiyik et al, 2012). During our investigation into the possible relationship between glucose repression and the oxidative stress response, we made the unexpected discovery that these cells display the same characteristics as diabetic cells in response to glucose since the regulatory mechanisms of glucose usage are highly conserved between S. pombe and human cells.…”

Section: Introductionmentioning

confidence: 99%

A potential protective role for thiamine in glucose-driven oxidative stress

Palabıyık¹,

Ghods²,

Uçar³

2014

Genet. Mol. Res.

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ABSTRACT.The relationship between glucose repression and the oxidative stress response was investigated in Schizosaccharomyces pombe wild type cells (972h -) and glucose repression resistant mutant type cells (ird11). We aimed to reveal the mechanism of simultaneous resistance to glucose repression and oxidative stress in ird11 mutants. Compared to the wild type, the expression of the sty1 gene was not altered in the ird11 mutant under normal growth conditions, but decreased after exposure to H 2 O 2 . This effect was clearly explained by the immunoblotting results, which showed elevated levels of a much more stable phosphorylated form of Sty1 mitogen-activated protein kinase in the ird11 mutant. Increased ght3 gene expression levels were also found, which may play a role in protecting the ird11 mutant from the deleterious effects of oxidative stress. In addition, decreased expression levels of glycolytic enzyme enolase-and thiamine synthesis/ transport-related genes were detected. This might have resulted from the flux redirection toward mitochondrial respiration, which would enhance NADPH generation to prevent the high reactive oxygen species accumulation that is generated by respiration. Some evidence supported a flux shift toward fermentation as well as respiration. We conclude that a defect in the glucose-sensing signaling pathway in Effects of thiamine in oxidative stress ird11 mutants likely causes erroneous low glucose-sensing signaling and high ATP production. This most likely occurs because high glucose availability in the medium induces an impairment in the respiratory chain and fermentation balance in these cells, which might explain the glucose repression and oxidative stress resistance in ird11 compared to the wild type.

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Isolation and Characterization of Glucose Derepressed Invertase Mutants fromSchizosaccharomyces pombe

Cited by 20 publications

References 15 publications

Effects of glucose sensing/signaling on oxidative stress response in glucose repression mutants of Schizosaccharomyces pombe

Effects of glucose sensing/signaling on oxidative stress response in glucose repression mutants of Schizosaccharomyces pombe

Characterization of two different types of UDP-glucose/-galactose4-epimerase involved in galactosylation in fission yeast

A potential protective role for thiamine in glucose-driven oxidative stress

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