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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 associate with their target genes upon switching of the carbon source from glucose to maltose in a manner dependent on each other.(A) Atf1 and Pcr1 associate with agl1 in maltose-utilizing cells. Cells expressing the indicated proteins were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins at the target regions of agl1 (presented in Figure 3A) were normalized against their levels at act1. Glc: glucose; Mal: maltose. *P<0.05, **P<0.01 (Student's t-test). (B) The increased level of Atf1 at agl1 depends upon Pcr1, and vice versa. Cells of the indicated strains were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins associated with the indicated genes relative to the levels associated with act1 in cells grown in maltose-containing medium. The fold change compared to levels in cells grown in glucose-containing medium is shown. Error bars, s.d. (n=3).
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pone-0080572-g004: Atf1 and Pcr1 associate with their target genes upon switching of the carbon source from glucose to maltose in a manner dependent on each other.(A) Atf1 and Pcr1 associate with agl1 in maltose-utilizing cells. Cells expressing the indicated proteins were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins at the target regions of agl1 (presented in Figure 3A) were normalized against their levels at act1. Glc: glucose; Mal: maltose. *P<0.05, **P<0.01 (Student's t-test). (B) The increased level of Atf1 at agl1 depends upon Pcr1, and vice versa. Cells of the indicated strains were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins associated with the indicated genes relative to the levels associated with act1 in cells grown in maltose-containing medium. The fold change compared to levels in cells grown in glucose-containing medium is shown. Error bars, s.d. (n=3).

Mentions: The in vivo binding sites of Atf1 and Pcr1 are degenerate in sequence and do not always match the TGACGT motif [14]. By surveying the degenerate sequences, potential binding sites of Atf1 and Pcr1 were identified interspersed in the promoter and coding region of agl1 (Table S4). Therefore, we next analyzed the distribution patterns of Atf1 and Pcr1 around the agl1 gene before and after the carbon source was changed. Enrichment of Atf1 and Pcr1 at the pdr1 gene, which is downstream of agl1, did not change after the carbon source was changed (5 kb in Figure 4A). By contrast, Atf1 and Pcr1 associated preferentially with the promoter, and to a lesser extent with the coding region, of agl1 after the carbon source was changed from glucose to maltose (Figure 4A). The associations of Atf1 and Pcr1 with the coding regions of fbp1 and gpx1 also increased when the carbon source was changed (Figure 4B), suggesting that Atf1 and Pcr1 associate with the coding regions of their target genes. The increased enrichment of Atf1 at its target genes was abolished when Pcr1 was depleted, and vice versa (Figure 4B), indicating that these two transcription factors act as a heteromer. These observations also strongly suggest that Atf1-Pcr1 heteromers associate with the promoters and coding regions of target genes, including agl1, in response to environmental changes.


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 associate with their target genes upon switching of the carbon source from glucose to maltose in a manner dependent on each other.(A) Atf1 and Pcr1 associate with agl1 in maltose-utilizing cells. Cells expressing the indicated proteins were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins at the target regions of agl1 (presented in Figure 3A) were normalized against their levels at act1. Glc: glucose; Mal: maltose. *P<0.05, **P<0.01 (Student's t-test). (B) The increased level of Atf1 at agl1 depends upon Pcr1, and vice versa. Cells of the indicated strains were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins associated with the indicated genes relative to the levels associated with act1 in cells grown in maltose-containing medium. The fold change compared to levels in cells grown in glucose-containing medium is shown. Error bars, s.d. (n=3).
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Related In: Results  -  Collection

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pone-0080572-g004: Atf1 and Pcr1 associate with their target genes upon switching of the carbon source from glucose to maltose in a manner dependent on each other.(A) Atf1 and Pcr1 associate with agl1 in maltose-utilizing cells. Cells expressing the indicated proteins were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins at the target regions of agl1 (presented in Figure 3A) were normalized against their levels at act1. Glc: glucose; Mal: maltose. *P<0.05, **P<0.01 (Student's t-test). (B) The increased level of Atf1 at agl1 depends upon Pcr1, and vice versa. Cells of the indicated strains were cultured as described in Figure 3B and used for ChIP analysis. The levels of the indicated proteins associated with the indicated genes relative to the levels associated with act1 in cells grown in maltose-containing medium. The fold change compared to levels in cells grown in glucose-containing medium is shown. Error bars, s.d. (n=3).
Mentions: The in vivo binding sites of Atf1 and Pcr1 are degenerate in sequence and do not always match the TGACGT motif [14]. By surveying the degenerate sequences, potential binding sites of Atf1 and Pcr1 were identified interspersed in the promoter and coding region of agl1 (Table S4). Therefore, we next analyzed the distribution patterns of Atf1 and Pcr1 around the agl1 gene before and after the carbon source was changed. Enrichment of Atf1 and Pcr1 at the pdr1 gene, which is downstream of agl1, did not change after the carbon source was changed (5 kb in Figure 4A). By contrast, Atf1 and Pcr1 associated preferentially with the promoter, and to a lesser extent with the coding region, of agl1 after the carbon source was changed from glucose to maltose (Figure 4A). The associations of Atf1 and Pcr1 with the coding regions of fbp1 and gpx1 also increased when the carbon source was changed (Figure 4B), suggesting that Atf1 and Pcr1 associate with the coding regions of their target genes. The increased enrichment of Atf1 at its target genes was abolished when Pcr1 was depleted, and vice versa (Figure 4B), indicating that these two transcription factors act as a heteromer. These observations also strongly suggest that Atf1-Pcr1 heteromers associate with the promoters and coding regions of target genes, including agl1, in response to environmental changes.

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