AUR1, a novel gene conferring aureobasidin resistance onSaccharomyces cerevisiae: a study of defective morphologies in Aur1p-depleted cells

Hashida-Okado, Takashi; Ogawa, Atsuko; Endo, Masahiro; Yasumoto, Ryoma; Takesako, Kazutoh; Kato, Ikunoshin

doi:10.1007/bf02172923

Cited by 69 publications

(82 citation statements)

References 21 publications

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“…Thus, replacement of the YAP1 gene or PGK promoter with the wild-type loci by homologous recombination will not confer the resistance phenotype to the recombinant cell. This situation is different from drug-resistance markers made from yeast genes with point mutations such as SMR1-410, cyh2, ARO4-OFP, AUR1-C, LEU4-1, FZF1-4 and PDR3-9 (Casey et al, 1988;Del Pozo et al, 1991;Shimura et al, 1993;Hashida-Okado et al, 1996;Bendoni et al, 1999;Park et al, 1999;Locková and Subík, 1999). The point mutations in the mutant genes could be replaced by homologous recombination with their respective wild-type genes when transformed.…”

Section: Discussionmentioning

confidence: 96%

“…The promoter of the highly expressed and constitutive yeast gene PGK (Kingsman et al, 1990) was fused to YAP1 to overexpress the gene product. Overexpression of YAP1 conferred cells with resistance to cerulenin, a fatty acid synthesis inhibitor (Kawaguchi et al, 1982) and also to cycloheximide, a protein synthesis inhibitor, but not to other drugs, including aureobasidin A (Hashida-Okado et al, 1996), nystatin (Snow, 1966) and amphotericin B (Woods and Ahmed, 1968). The multiple and specific drug resistance of PGKp-YAP1 was used to select PGKp-YAP1 transformants from a background of resistant cells.…”

Section: Introductionmentioning

confidence: 99%

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Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae

Akada

Shimizu

Matsushita

et al. 2001

Yeast

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Drug-resistance markers for yeast transformation are useful because they can be applied to strains without auxotrophic mutations. However, they are susceptible to technical difficulties, namely lower transformation efficiency and the appearance of drug-resistant mutants without the marker. To avoid these problems, we have constructed a phosphoglycerate kinase (PGK) promoter-driven YAP1 expression cassette, called PGKp-YAP1. Yeast cells containing PGKp-YAP1 were resistant to cycloheximide, a protein synthesis inhibitor, and also to cerulenin, a fatty acid synthesis inhibitor, but not to other drugs tested. The transformation efficiency of PGKp-YAP1 using cerulenin selection was comparable to that using a URA3 auxotrophic marker when low concentrations of cerulenin were used. Non-transformed drug-resistant colonies did appear on the low-concentration cerulenin plates. However, these non-transformed colonies could easily be identified, based on their cycloheximide sensitivity and/or their resistance to aureobasidin A to which the transformants were sensitive. Therefore, the dual drug resistance of PGKp-YAP1 could be used as an effective selection for PGKp-YAP1 recipient cells. The PGKp-YAP1 marker was used to disrupt the LYS2 gene and to transform an industrial yeast strain, indicating that this marker can be used for efficient and reliable gene manipulations in any Saccharomyces cerevisiae strain.

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Section: Discussionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae

Akada

Shimizu

Matsushita

et al. 2001

Yeast

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“…(Hashida-Okado et al, 1996), and Aspergillus nidulans (G275V) (Kuroda et al, 1999). The L197 of the AUR1 protein is highly conserved among B. cinerea, S. cerevisiae, A. nidulans and A. fumigatus (Fig.…”

Section: Isolation and Characterization Of The Aur1 Gene From Aba-resmentioning

confidence: 99%

“…The transfer of the phosphorylinositol group from phosphatidylinositol (PI) to ceramide is the key step, which was catalyzed by IPC synthase in fungi (Dickson and Lester, 2002;Dickson et al, 2006;Obeid et al, 2002). IPC synthase encoded by the AUR1 gene in yeast is a recognized target for antifungals (Hashida-Okado et al, 1996;Kuroda et al, 1999;Nagiec et al, 1997). The IPC synthase gene (AUR1) has been identified in various fungal species (Aeed et al, 2009;Heidler and Radding, 1995;Kuroda et al, 1999;Nagiec et al, 1997;Zhong et al, 2000).…”

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confidence: 99%

“…The fungal cells treated with AbA or the inhibition of AUR1 expression can cause pleiotropic morphological changes, including the disappearance of microtubules, degradation of tubulin and abnormal deposition of chitin, ultimately leading to cell death (Cerantola et al, 2009;Cheng et al, 2001;Hashida-Okado et al, 1996). Studies involving drug resistance in fungi and protozoa have shown that mutations in the AUR1 confer resistance to the antifungal drug AbA, and mutants can resist the inhibition of AbA against IPC synthase activity (Hashida- Okado et al, 1996;Heidler and Radding, 2000;Kuroda et al, 1999;Nagiec et al, 1997;Zhong et al, 2000). Many phytopathogens can secrete oxalic acid which is the essential pathogenic determinant involved in disease development.…”

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confidence: 99%

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Characteristics of inositol phosphorylceramide synthase and effects of aureobasidin A on growth and pathogenicity of <i>Botrytis cinerea</i>

Wang

Guo

et al. 2015

J. Gen. Appl. Microbiol.

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IntroductionSphingolipids are known to be important components Inositol phosphorylceramide (IPC) synthase is the key enzyme with highly conserved sequences, which is involved in fungal sphingolipid biosynthesis. The antibiotic aureobasidin A (AbA) induces the death of fungi through inhibiting IPC synthase activity. The mutations of AUR1 gene coding IPC synthase in fungi and protozoa causes a resistance to AbA. However, the mechanism of AbA resistance is still elusive. In this paper, we generated two mutants of Botrytis cinerea with AbA-resistance, BcAUR1a and BcAUR1b, through UV irradiation. BcAUR1a lost an intron and BcAUR1b had three amino acid mutations (L197P, F288S and T323A) in the AUR1 gene. AbA strongly inhibits the activity of IPC synthase in wild-type B. cinerea, which leads to distinct changes in cell morphology, including the delay in conidial germination, excessive branching near the tip of the germ tube and mycelium, and the inhibition of the mycelium growth. Further, AbA prevents the infection of wild-type B. cinerea in tomato fruits via reducing oxalic acid secretion and the activity of cellulase and pectinase. On the contrary, AbA has no effect on the growth and pathogenicity of the two mutants. Although both mutants show a similar AbA resistance, the molecular mechanisms might be different between the two mutants.

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A positive selection for plasmid loss inSaccharomyces cerevisiae using galactose-inducible growth inhibitory sequences

Kawahata

Amari

Akada

1999

Yeast

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AUR1, a novel gene conferring aureobasidin resistance onSaccharomyces cerevisiae: a study of defective morphologies in Aur1p-depleted cells

Cited by 69 publications

References 21 publications

Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae

Use of a YAP1 overexpression cassette conferring specific resistance to cerulenin and cycloheximide as an efficient selectable marker in the yeast Saccharomyces cerevisiae

Characteristics of inositol phosphorylceramide synthase and effects of aureobasidin A on growth and pathogenicity of <i>Botrytis cinerea</i>

A positive selection for plasmid loss inSaccharomyces cerevisiae using galactose-inducible growth inhibitory sequences

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