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The transcription factors Atf1 and Pcr1 are essential for transcriptional induction of the extracellular maltase Agl1 in fission yeast.

Kato H, Kira S, Kawamukai M - PLoS ONE (2013)

Bottom Line: When the carbon source was switched from glucose to maltose, Atf1 and Pcr1 associated with the promoters and coding regions of agl1, fbp1, and gpx1, indicating that the Atf1-Pcr1 heteromer binds a variety of regions in its target genes to induce their transcription.In addition, the association of Mediator with these genes was dependent on Atf1 and Pcr1.These data indicate that Atf1 and Pcr1 induce the transcription of agl1, which allows efficient utilization of extracellular maltose.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan ; Department of Biochemistry, Shimane University School of Medicine, Izumo, Japan ; PRESTO, Japan Science and Technology Agency (JST), Saitama, Japan.

ABSTRACT
The fission yeast Schizosaccharomyces pombe secretes the extracellular maltase Agl1, which hydrolyzes maltose into glucose, thereby utilizing maltose as a carbon source. Whether other maltases contribute to efficient utilization of maltose and how Agl1 expression is regulated in response to switching of carbon sources are unknown. In this study, we show that three other possible maltases and the maltose transporter Sut1 are not required for efficient utilization of maltose. Transcription of agl1 was induced when the carbon source was changed from glucose to maltose. This was dependent on Atf1 and Pcr1, which are highly conserved transcription factors that regulate stress-responsive genes in various stress conditions. Atf1 and Pcr1 generally bind the TGACGT motif as a heterodimer. The agl1 gene lacks the exact motif, but has many degenerate TGACGT motifs in its promoter and coding region. When the carbon source was switched from glucose to maltose, Atf1 and Pcr1 associated with the promoters and coding regions of agl1, fbp1, and gpx1, indicating that the Atf1-Pcr1 heteromer binds a variety of regions in its target genes to induce their transcription. In addition, the association of Mediator with these genes was dependent on Atf1 and Pcr1. These data indicate that Atf1 and Pcr1 induce the transcription of agl1, which allows efficient utilization of extracellular maltose.

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Atf1 and Pcr1 are required for efficient maltose utilization.(A) Fission yeast spores fail to form colonies when grown on maltose-containing medium. Wild-type (mating-type: h90) tetrads were dissected on YE (glucose-containing) and YEM (maltose-containing) plates, and incubated at 30°C for 3 days. (B) atf1∆ and pcr1∆ mutants grow slowly on maltose-containing plates. Wild-type, atf1∆, and pcr1∆ strains were streaked on plates containing the indicated sugar(s) and incubated at 30°C for 3 days. (C) Growth of atf1∆ and pcr1∆ mutants does not recover following switching from glucose- to maltose-containing medium. Cells of the indicated strains exponentially growing in YE were washed twice, resuspended in YE or YEM, and incubated at 30°C for the indicated number of hours.
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pone-0080572-g001: Atf1 and Pcr1 are required for efficient maltose utilization.(A) Fission yeast spores fail to form colonies when grown on maltose-containing medium. Wild-type (mating-type: h90) tetrads were dissected on YE (glucose-containing) and YEM (maltose-containing) plates, and incubated at 30°C for 3 days. (B) atf1∆ and pcr1∆ mutants grow slowly on maltose-containing plates. Wild-type, atf1∆, and pcr1∆ strains were streaked on plates containing the indicated sugar(s) and incubated at 30°C for 3 days. (C) Growth of atf1∆ and pcr1∆ mutants does not recover following switching from glucose- to maltose-containing medium. Cells of the indicated strains exponentially growing in YE were washed twice, resuspended in YE or YEM, and incubated at 30°C for the indicated number of hours.

Mentions: In a routine genetic analysis of fission yeast, we noticed that spores did not germinate on plates that contained 3% maltose instead of 3% glucose (Figure 1A), indicating that spores cannot utilize maltose. We identified atf1 and pcr1 in a reverse genetic screen of genes that contribute to maltose hydrolysis. Although atf1∆ and pcr1∆ cells grow slightly slower than wild-type cells on YE plates that contain glucose as a carbon source [11,32], atf1∆ cells grew markedly slower than wild-type cells on YEM plates (Figure 1B). pcr1∆ cells also grew slower than wild-type cells on YEM plates, but only when the pcr1∆ and wild-type cells were well-separated from each other (Figure 1B). atf1∆ and pcr1∆ cells did not show a severe growth defect when grown on a plate containing both maltose and glucose (Figure 1B), indicating that maltose is not toxic to these mutants and that Atf1 and Pcr1 are required for efficient utilization of maltose. Wild-type cells exponentially growing in YE medium at 30°C temporarily stopped growing when the carbon source was changed from glucose to maltose (Figure 1C). About 10 hours after this switch, the cells resumed growing. The timing of this growth recovery was dependent on the density of cells when the carbon source was switched (data not shown), suggesting that growth resumes when the cells have secreted a sufficient amount of maltase. atf1∆ and pcr1∆ mutants also stopped growing when the carbon source was changed; however, unlike wild-type cells, they did not resume growing (Figure 1C). These data indicate that Atf1 and Pcr1 are required for efficient maltose utilization.


The transcription factors Atf1 and Pcr1 are essential for transcriptional induction of the extracellular maltase Agl1 in fission yeast.

Kato H, Kira S, Kawamukai M - PLoS ONE (2013)

Atf1 and Pcr1 are required for efficient maltose utilization.(A) Fission yeast spores fail to form colonies when grown on maltose-containing medium. Wild-type (mating-type: h90) tetrads were dissected on YE (glucose-containing) and YEM (maltose-containing) plates, and incubated at 30°C for 3 days. (B) atf1∆ and pcr1∆ mutants grow slowly on maltose-containing plates. Wild-type, atf1∆, and pcr1∆ strains were streaked on plates containing the indicated sugar(s) and incubated at 30°C for 3 days. (C) Growth of atf1∆ and pcr1∆ mutants does not recover following switching from glucose- to maltose-containing medium. Cells of the indicated strains exponentially growing in YE were washed twice, resuspended in YE or YEM, and incubated at 30°C for the indicated number of hours.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3818258&req=5

pone-0080572-g001: Atf1 and Pcr1 are required for efficient maltose utilization.(A) Fission yeast spores fail to form colonies when grown on maltose-containing medium. Wild-type (mating-type: h90) tetrads were dissected on YE (glucose-containing) and YEM (maltose-containing) plates, and incubated at 30°C for 3 days. (B) atf1∆ and pcr1∆ mutants grow slowly on maltose-containing plates. Wild-type, atf1∆, and pcr1∆ strains were streaked on plates containing the indicated sugar(s) and incubated at 30°C for 3 days. (C) Growth of atf1∆ and pcr1∆ mutants does not recover following switching from glucose- to maltose-containing medium. Cells of the indicated strains exponentially growing in YE were washed twice, resuspended in YE or YEM, and incubated at 30°C for the indicated number of hours.
Mentions: In a routine genetic analysis of fission yeast, we noticed that spores did not germinate on plates that contained 3% maltose instead of 3% glucose (Figure 1A), indicating that spores cannot utilize maltose. We identified atf1 and pcr1 in a reverse genetic screen of genes that contribute to maltose hydrolysis. Although atf1∆ and pcr1∆ cells grow slightly slower than wild-type cells on YE plates that contain glucose as a carbon source [11,32], atf1∆ cells grew markedly slower than wild-type cells on YEM plates (Figure 1B). pcr1∆ cells also grew slower than wild-type cells on YEM plates, but only when the pcr1∆ and wild-type cells were well-separated from each other (Figure 1B). atf1∆ and pcr1∆ cells did not show a severe growth defect when grown on a plate containing both maltose and glucose (Figure 1B), indicating that maltose is not toxic to these mutants and that Atf1 and Pcr1 are required for efficient utilization of maltose. Wild-type cells exponentially growing in YE medium at 30°C temporarily stopped growing when the carbon source was changed from glucose to maltose (Figure 1C). About 10 hours after this switch, the cells resumed growing. The timing of this growth recovery was dependent on the density of cells when the carbon source was switched (data not shown), suggesting that growth resumes when the cells have secreted a sufficient amount of maltase. atf1∆ and pcr1∆ mutants also stopped growing when the carbon source was changed; however, unlike wild-type cells, they did not resume growing (Figure 1C). These data indicate that Atf1 and Pcr1 are required for efficient maltose utilization.

Bottom Line: When the carbon source was switched from glucose to maltose, Atf1 and Pcr1 associated with the promoters and coding regions of agl1, fbp1, and gpx1, indicating that the Atf1-Pcr1 heteromer binds a variety of regions in its target genes to induce their transcription.In addition, the association of Mediator with these genes was dependent on Atf1 and Pcr1.These data indicate that Atf1 and Pcr1 induce the transcription of agl1, which allows efficient utilization of extracellular maltose.

View Article: PubMed Central - PubMed

Affiliation: Department of Life Science and Biotechnology, Faculty of Life and Environmental Science, Shimane University, Matsue, Japan ; Department of Biochemistry, Shimane University School of Medicine, Izumo, Japan ; PRESTO, Japan Science and Technology Agency (JST), Saitama, Japan.

ABSTRACT
The fission yeast Schizosaccharomyces pombe secretes the extracellular maltase Agl1, which hydrolyzes maltose into glucose, thereby utilizing maltose as a carbon source. Whether other maltases contribute to efficient utilization of maltose and how Agl1 expression is regulated in response to switching of carbon sources are unknown. In this study, we show that three other possible maltases and the maltose transporter Sut1 are not required for efficient utilization of maltose. Transcription of agl1 was induced when the carbon source was changed from glucose to maltose. This was dependent on Atf1 and Pcr1, which are highly conserved transcription factors that regulate stress-responsive genes in various stress conditions. Atf1 and Pcr1 generally bind the TGACGT motif as a heterodimer. The agl1 gene lacks the exact motif, but has many degenerate TGACGT motifs in its promoter and coding region. When the carbon source was switched from glucose to maltose, Atf1 and Pcr1 associated with the promoters and coding regions of agl1, fbp1, and gpx1, indicating that the Atf1-Pcr1 heteromer binds a variety of regions in its target genes to induce their transcription. In addition, the association of Mediator with these genes was dependent on Atf1 and Pcr1. These data indicate that Atf1 and Pcr1 induce the transcription of agl1, which allows efficient utilization of extracellular maltose.

Show MeSH
Related in: MedlinePlus