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Regulation of wee1(+) expression during meiosis in fission yeast.

Murakami-Tonami Y, Ohtsuka H, Aiba H, Murakami H - Cell Cycle (2014)

Bottom Line: Conversely, deletion of Mei4p did not decrease expression of wee1(+) but inhibited that of the pseudogene.In addition, deletion of Mei4p-binding regions delayed repression of wee1(+) expression as well as induction of expression of the pseudogene.These results suggest that repression of wee1(+) expression is primarily owing to Mei4p-mediated transcriptional interference.

View Article: PubMed Central - PubMed

Affiliation: a Aichi Cancer Center Research Institute ; Division of Molecular Oncology ; Nagoya , Japan.

ABSTRACT
In eukaryotes, the cyclin-dependent kinase Cdk1p (Cdc2p) plays a central role in entry into and progression through nuclear division during mitosis and meiosis. Cdk1p is activated during meiotic nuclear divisions by dephosphorylation of its tyrosine-15 residue. The phosphorylation status of this residue is largely determined by the Wee1p kinase and the Cdc25p phosphatase. In fission yeast, the forkhead-type transcription factor Mei4p is essential for entry into the first meiotic nuclear division. We recently identified cdc25(+) as an essential target of Mei4p in the control of entry into meiosis I. Here, we show that wee1(+) is another important target of Mei4p in the control of entry into meiosis I. Mei4p bound to the upstream region of wee1(+) in vivo and in vitro and inhibited expression of wee1(+), whereas Mei4p positively regulated expression of the adjacent pseudogene. Overexpression of Mei4p inhibited expression of wee1(+) and induced that of the pseudogene. Conversely, deletion of Mei4p did not decrease expression of wee1(+) but inhibited that of the pseudogene. In addition, deletion of Mei4p-binding regions delayed repression of wee1(+) expression as well as induction of expression of the pseudogene. These results suggest that repression of wee1(+) expression is primarily owing to Mei4p-mediated transcriptional interference.

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Effects of deletion of the FLEX sites near to wee1+ on wee1+ gene expression. (A) Schematic representation of the wee1+ and SPCC18B5.02c loci according to the Sanger Center database. ORF, open reading frame; (B) The region of the wee1+ gene containing FLEX1, FLEX2, FLEX3, and FLEX4 was deleted (HM5678). Meiosis was induced in the resulting cells as described in Fig. 1. Samples were collected at the indicated time points after meiosis induction and subjected to Northern blot analysis of wee1+ and SPCC18B5.02c mRNA. Ribosomal RNA (rRNA) was stained with ethidium bromide as the loading control.
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f0003: Effects of deletion of the FLEX sites near to wee1+ on wee1+ gene expression. (A) Schematic representation of the wee1+ and SPCC18B5.02c loci according to the Sanger Center database. ORF, open reading frame; (B) The region of the wee1+ gene containing FLEX1, FLEX2, FLEX3, and FLEX4 was deleted (HM5678). Meiosis was induced in the resulting cells as described in Fig. 1. Samples were collected at the indicated time points after meiosis induction and subjected to Northern blot analysis of wee1+ and SPCC18B5.02c mRNA. Ribosomal RNA (rRNA) was stained with ethidium bromide as the loading control.

Mentions: We next examined whether the FLEX sequences associated with wee1+ function as transcriptional cis elements. We deleted the chromosomal region containing FLEX1, FLEX2, FLEX3, and FLEX4 of wee1+ (Fig. 3A). In the resulting cells, wee1+ mRNA was downregulated 1 h later than in WT cells following the induction of meiosis, and induction of the adjacent pseudogene was also delayed (Fig. 3B). This result shows that 4 of the FLEX elements located close to wee1+ are required for efficient down-regulation of wee1+ mRNA and up-regulation of the adjacent pseudogene mRNA. This suggests that these sites serve as cis-acting elements for Mei4p.Figure 3.


Regulation of wee1(+) expression during meiosis in fission yeast.

Murakami-Tonami Y, Ohtsuka H, Aiba H, Murakami H - Cell Cycle (2014)

Effects of deletion of the FLEX sites near to wee1+ on wee1+ gene expression. (A) Schematic representation of the wee1+ and SPCC18B5.02c loci according to the Sanger Center database. ORF, open reading frame; (B) The region of the wee1+ gene containing FLEX1, FLEX2, FLEX3, and FLEX4 was deleted (HM5678). Meiosis was induced in the resulting cells as described in Fig. 1. Samples were collected at the indicated time points after meiosis induction and subjected to Northern blot analysis of wee1+ and SPCC18B5.02c mRNA. Ribosomal RNA (rRNA) was stained with ethidium bromide as the loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612672&req=5

f0003: Effects of deletion of the FLEX sites near to wee1+ on wee1+ gene expression. (A) Schematic representation of the wee1+ and SPCC18B5.02c loci according to the Sanger Center database. ORF, open reading frame; (B) The region of the wee1+ gene containing FLEX1, FLEX2, FLEX3, and FLEX4 was deleted (HM5678). Meiosis was induced in the resulting cells as described in Fig. 1. Samples were collected at the indicated time points after meiosis induction and subjected to Northern blot analysis of wee1+ and SPCC18B5.02c mRNA. Ribosomal RNA (rRNA) was stained with ethidium bromide as the loading control.
Mentions: We next examined whether the FLEX sequences associated with wee1+ function as transcriptional cis elements. We deleted the chromosomal region containing FLEX1, FLEX2, FLEX3, and FLEX4 of wee1+ (Fig. 3A). In the resulting cells, wee1+ mRNA was downregulated 1 h later than in WT cells following the induction of meiosis, and induction of the adjacent pseudogene was also delayed (Fig. 3B). This result shows that 4 of the FLEX elements located close to wee1+ are required for efficient down-regulation of wee1+ mRNA and up-regulation of the adjacent pseudogene mRNA. This suggests that these sites serve as cis-acting elements for Mei4p.Figure 3.

Bottom Line: Conversely, deletion of Mei4p did not decrease expression of wee1(+) but inhibited that of the pseudogene.In addition, deletion of Mei4p-binding regions delayed repression of wee1(+) expression as well as induction of expression of the pseudogene.These results suggest that repression of wee1(+) expression is primarily owing to Mei4p-mediated transcriptional interference.

View Article: PubMed Central - PubMed

Affiliation: a Aichi Cancer Center Research Institute ; Division of Molecular Oncology ; Nagoya , Japan.

ABSTRACT
In eukaryotes, the cyclin-dependent kinase Cdk1p (Cdc2p) plays a central role in entry into and progression through nuclear division during mitosis and meiosis. Cdk1p is activated during meiotic nuclear divisions by dephosphorylation of its tyrosine-15 residue. The phosphorylation status of this residue is largely determined by the Wee1p kinase and the Cdc25p phosphatase. In fission yeast, the forkhead-type transcription factor Mei4p is essential for entry into the first meiotic nuclear division. We recently identified cdc25(+) as an essential target of Mei4p in the control of entry into meiosis I. Here, we show that wee1(+) is another important target of Mei4p in the control of entry into meiosis I. Mei4p bound to the upstream region of wee1(+) in vivo and in vitro and inhibited expression of wee1(+), whereas Mei4p positively regulated expression of the adjacent pseudogene. Overexpression of Mei4p inhibited expression of wee1(+) and induced that of the pseudogene. Conversely, deletion of Mei4p did not decrease expression of wee1(+) but inhibited that of the pseudogene. In addition, deletion of Mei4p-binding regions delayed repression of wee1(+) expression as well as induction of expression of the pseudogene. These results suggest that repression of wee1(+) expression is primarily owing to Mei4p-mediated transcriptional interference.

Show MeSH
Related in: MedlinePlus