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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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Phosphorylation of Cmk1 in response to osmotic stress. (A) cmk1:HA strain was treated with 100 mM CaCl2 or 1M KCl when indicated and Cmk1 phosphorylation was detected by Western blot with anti-HA antibodies (top). (B) Time course of cells harbouring endogenous cmk1:HA gene exposed to 1M KCl. Cmk1 protein level was analysed at times indicated as in A (top). (C) The same strain cmk1:HA was treated with 1M KCl and 20 mM EGTA when indicated and Cmk1 was detected as in A (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).
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Figure 2: Phosphorylation of Cmk1 in response to osmotic stress. (A) cmk1:HA strain was treated with 100 mM CaCl2 or 1M KCl when indicated and Cmk1 phosphorylation was detected by Western blot with anti-HA antibodies (top). (B) Time course of cells harbouring endogenous cmk1:HA gene exposed to 1M KCl. Cmk1 protein level was analysed at times indicated as in A (top). (C) The same strain cmk1:HA was treated with 1M KCl and 20 mM EGTA when indicated and Cmk1 was detected as in A (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).

Mentions: An increase in extracellular Ca2+ can also provoke osmotic stress in cells; therefore, we also analysed the Cmk1 protein in response to osmotic stress by KCl. The Cmk1 protein was also phosphorylated in the presence of KCl, displaying a slow migration band as in the response to Ca2+ (Figure 2A). Cmk1 phosphorylation was concomitant with KCl exposure, as shown in the time course of Figure 2B.


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)

Phosphorylation of Cmk1 in response to osmotic stress. (A) cmk1:HA strain was treated with 100 mM CaCl2 or 1M KCl when indicated and Cmk1 phosphorylation was detected by Western blot with anti-HA antibodies (top). (B) Time course of cells harbouring endogenous cmk1:HA gene exposed to 1M KCl. Cmk1 protein level was analysed at times indicated as in A (top). (C) The same strain cmk1:HA was treated with 1M KCl and 20 mM EGTA when indicated and Cmk1 was detected as in A (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 2: Phosphorylation of Cmk1 in response to osmotic stress. (A) cmk1:HA strain was treated with 100 mM CaCl2 or 1M KCl when indicated and Cmk1 phosphorylation was detected by Western blot with anti-HA antibodies (top). (B) Time course of cells harbouring endogenous cmk1:HA gene exposed to 1M KCl. Cmk1 protein level was analysed at times indicated as in A (top). (C) The same strain cmk1:HA was treated with 1M KCl and 20 mM EGTA when indicated and Cmk1 was detected as in A (top). Anti-PSTAIR antibodies were used to probe Cdc2 as a loading control (bottom).
Mentions: An increase in extracellular Ca2+ can also provoke osmotic stress in cells; therefore, we also analysed the Cmk1 protein in response to osmotic stress by KCl. The Cmk1 protein was also phosphorylated in the presence of KCl, displaying a slow migration band as in the response to Ca2+ (Figure 2A). Cmk1 phosphorylation was concomitant with KCl exposure, as shown in the time course of Figure 2B.

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