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Negative feedback regulation of calcineurin-dependent Prz1 transcription factor by the CaMKK-CaMK1 axis in fission yeast.

Cisneros-Barroso E, Yance-Chávez T, Kito A, Sugiura R, Gómez-Hierro A, Giménez-Zaragoza D, Aligue R - Nucleic Acids Res. (2014)

Bottom Line: Then, active Cmk1 binds, phosphorylates and inactivates Prz1 transcription activity whilst at the same time cmk1 expression is enhanced by Prz1 in response to Ca(2+).This work reveals that Cmk1 kinase activated by the newly identified Ckk2 counteracts calcineurin function by negatively regulating Prz1 activity which in turn is involved in activating cmk1 gene transcription.These results are the first insights into Cmk1 and Ckk2 function in Schizosaccharomyces pombe.

View Article: PubMed Central - PubMed

Affiliation: Departament de Biologia Cellular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, Institute of Biomedical Research August Pi i Sunyer (IDIBAPS), Barcelona 08036, Catalunya, Spain.

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Transcription of Cmk1 is regulated by Prz1 in response to Ca2+. (A) Promoter analysis of the cmk1 gene. Calcineurin-dependent response element (CDRE), AGCCTC/GAGGCT is boxed and putative Ca2+-dependent response elements CAACT/AGTTG are boxed in bold. (B) Cmk1 gene expression was analysed in wild-type (wt) and prz1 deleted (Δprz1) cells treated with 100 mM CaCl2 at different times. The mRNA of the cells was extracted and the cmk1 mRNA level was analysed by RT-PCR using the oligonucleotides Cmk1 RT Fw and Rv. (C) Cmk1 protein level was analysed from the same cells as in B by Western blot with anti-HA antibodies (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).
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Figure 3: Transcription of Cmk1 is regulated by Prz1 in response to Ca2+. (A) Promoter analysis of the cmk1 gene. Calcineurin-dependent response element (CDRE), AGCCTC/GAGGCT is boxed and putative Ca2+-dependent response elements CAACT/AGTTG are boxed in bold. (B) Cmk1 gene expression was analysed in wild-type (wt) and prz1 deleted (Δprz1) cells treated with 100 mM CaCl2 at different times. The mRNA of the cells was extracted and the cmk1 mRNA level was analysed by RT-PCR using the oligonucleotides Cmk1 RT Fw and Rv. (C) Cmk1 protein level was analysed from the same cells as in B by Western blot with anti-HA antibodies (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).

Mentions: An examination of the cmk1 promoter revealed the presence of a number of putative Ca2+-dependent response elements: CAACT/AGTTG located at 431–427, 300–296 and 273–269 bp upstream of ATG (Figure 3A), suggesting that Ca2+-induced expression of Cmk1 in fission yeast is the result of up-regulated transcription (19). In addition to the Ca2+-responsive elements, the cmk1 promoter also contains the previously described calcineurin-dependent response element (CDRE) AGCCTC/GAGGCT motif (35). This motif is found at the 296–291 position upstream of ATG in the cmk1 promoter (Figure 3A).


Negative feedback regulation of calcineurin-dependent Prz1 transcription factor by the CaMKK-CaMK1 axis in fission yeast.

Cisneros-Barroso E, Yance-Chávez T, Kito A, Sugiura R, Gómez-Hierro A, Giménez-Zaragoza D, Aligue R - Nucleic Acids Res. (2014)

Transcription of Cmk1 is regulated by Prz1 in response to Ca2+. (A) Promoter analysis of the cmk1 gene. Calcineurin-dependent response element (CDRE), AGCCTC/GAGGCT is boxed and putative Ca2+-dependent response elements CAACT/AGTTG are boxed in bold. (B) Cmk1 gene expression was analysed in wild-type (wt) and prz1 deleted (Δprz1) cells treated with 100 mM CaCl2 at different times. The mRNA of the cells was extracted and the cmk1 mRNA level was analysed by RT-PCR using the oligonucleotides Cmk1 RT Fw and Rv. (C) Cmk1 protein level was analysed from the same cells as in B by Western blot with anti-HA antibodies (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150787&req=5

Figure 3: Transcription of Cmk1 is regulated by Prz1 in response to Ca2+. (A) Promoter analysis of the cmk1 gene. Calcineurin-dependent response element (CDRE), AGCCTC/GAGGCT is boxed and putative Ca2+-dependent response elements CAACT/AGTTG are boxed in bold. (B) Cmk1 gene expression was analysed in wild-type (wt) and prz1 deleted (Δprz1) cells treated with 100 mM CaCl2 at different times. The mRNA of the cells was extracted and the cmk1 mRNA level was analysed by RT-PCR using the oligonucleotides Cmk1 RT Fw and Rv. (C) Cmk1 protein level was analysed from the same cells as in B by Western blot with anti-HA antibodies (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).
Mentions: An examination of the cmk1 promoter revealed the presence of a number of putative Ca2+-dependent response elements: CAACT/AGTTG located at 431–427, 300–296 and 273–269 bp upstream of ATG (Figure 3A), suggesting that Ca2+-induced expression of Cmk1 in fission yeast is the result of up-regulated transcription (19). In addition to the Ca2+-responsive elements, the cmk1 promoter also contains the previously described calcineurin-dependent response element (CDRE) AGCCTC/GAGGCT motif (35). This motif is found at the 296–291 position upstream of ATG in the cmk1 promoter (Figure 3A).

Bottom Line: Then, active Cmk1 binds, phosphorylates and inactivates Prz1 transcription activity whilst at the same time cmk1 expression is enhanced by Prz1 in response to Ca(2+).This work reveals that Cmk1 kinase activated by the newly identified Ckk2 counteracts calcineurin function by negatively regulating Prz1 activity which in turn is involved in activating cmk1 gene transcription.These results are the first insights into Cmk1 and Ckk2 function in Schizosaccharomyces pombe.

View Article: PubMed Central - PubMed

Affiliation: Departament de Biologia Cellular, Immunologia i Neurociències, Facultat de Medicina, Universitat de Barcelona, Institute of Biomedical Research August Pi i Sunyer (IDIBAPS), Barcelona 08036, Catalunya, Spain.

Show MeSH
Related in: MedlinePlus