Limits...
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.

Show MeSH

Related in: MedlinePlus

Ckk2 a new CaMKK in fission yeast. (A) Comparison of amino acid sequences of human and S. pombe CaMKK. Human CaMKK2 (CaMKK2), human CaMKK1 (CaMKK1), S. pombe ckk2 (Ckk2) and S. pombe ssp1 (Ssp1). Multiple sequences were aligned with Clustal Omega. (B) Ca2+ sensitivity. Wild-type (wt), Δcmk1, Δckk2 and Δcmk1 Δckk2 cells were grown in YES medium and spotted on YES plates containing different concentrations of CaCl2 and incubated for 3 days at 30°C. (C) Time course of wild-type (wt) and Δckk2 cells exposed to 100 mM CaCl2. The Cmk1 protein and phosphorylation mobility shift were analysed at the times indicated by Western blot using anti-HA antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom). (D) Time course of wild-type (wt), Δcmk1 and Δckk2 cells exposed to 100 mM CaCl2. The Cdc25 protein level was analysed at the times indicated by Western blot using anti-Cdc25 antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4150787&req=5

Figure 8: Ckk2 a new CaMKK in fission yeast. (A) Comparison of amino acid sequences of human and S. pombe CaMKK. Human CaMKK2 (CaMKK2), human CaMKK1 (CaMKK1), S. pombe ckk2 (Ckk2) and S. pombe ssp1 (Ssp1). Multiple sequences were aligned with Clustal Omega. (B) Ca2+ sensitivity. Wild-type (wt), Δcmk1, Δckk2 and Δcmk1 Δckk2 cells were grown in YES medium and spotted on YES plates containing different concentrations of CaCl2 and incubated for 3 days at 30°C. (C) Time course of wild-type (wt) and Δckk2 cells exposed to 100 mM CaCl2. The Cmk1 protein and phosphorylation mobility shift were analysed at the times indicated by Western blot using anti-HA antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom). (D) Time course of wild-type (wt), Δcmk1 and Δckk2 cells exposed to 100 mM CaCl2. The Cdc25 protein level was analysed at the times indicated by Western blot using anti-Cdc25 antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom).

Mentions: In addition to Ssp1, we found a second sequence homologous to mammalian CaMKK: SPC1919.01, named Ppk34 for the uncharacterised putative protein kinase 34 in a systematic deletion analysis of fission yeast kinases (45). Due to its homology to the CaMKK proteins, we have renamed it Ckk2. Ckk2 is most homologous to mammalian CaMKK2 (34%), followed by Ssp1 (32%) and CaMKK1 (28%) (Figure 8A and Table 1).


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)

Ckk2 a new CaMKK in fission yeast. (A) Comparison of amino acid sequences of human and S. pombe CaMKK. Human CaMKK2 (CaMKK2), human CaMKK1 (CaMKK1), S. pombe ckk2 (Ckk2) and S. pombe ssp1 (Ssp1). Multiple sequences were aligned with Clustal Omega. (B) Ca2+ sensitivity. Wild-type (wt), Δcmk1, Δckk2 and Δcmk1 Δckk2 cells were grown in YES medium and spotted on YES plates containing different concentrations of CaCl2 and incubated for 3 days at 30°C. (C) Time course of wild-type (wt) and Δckk2 cells exposed to 100 mM CaCl2. The Cmk1 protein and phosphorylation mobility shift were analysed at the times indicated by Western blot using anti-HA antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom). (D) Time course of wild-type (wt), Δcmk1 and Δckk2 cells exposed to 100 mM CaCl2. The Cdc25 protein level was analysed at the times indicated by Western blot using anti-Cdc25 antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom).
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 8: Ckk2 a new CaMKK in fission yeast. (A) Comparison of amino acid sequences of human and S. pombe CaMKK. Human CaMKK2 (CaMKK2), human CaMKK1 (CaMKK1), S. pombe ckk2 (Ckk2) and S. pombe ssp1 (Ssp1). Multiple sequences were aligned with Clustal Omega. (B) Ca2+ sensitivity. Wild-type (wt), Δcmk1, Δckk2 and Δcmk1 Δckk2 cells were grown in YES medium and spotted on YES plates containing different concentrations of CaCl2 and incubated for 3 days at 30°C. (C) Time course of wild-type (wt) and Δckk2 cells exposed to 100 mM CaCl2. The Cmk1 protein and phosphorylation mobility shift were analysed at the times indicated by Western blot using anti-HA antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom). (D) Time course of wild-type (wt), Δcmk1 and Δckk2 cells exposed to 100 mM CaCl2. The Cdc25 protein level was analysed at the times indicated by Western blot using anti-Cdc25 antibodies (top) and Cdc2 was probed as a loading control with anti-PSTAIR antibodies (bottom).
Mentions: In addition to Ssp1, we found a second sequence homologous to mammalian CaMKK: SPC1919.01, named Ppk34 for the uncharacterised putative protein kinase 34 in a systematic deletion analysis of fission yeast kinases (45). Due to its homology to the CaMKK proteins, we have renamed it Ckk2. Ckk2 is most homologous to mammalian CaMKK2 (34%), followed by Ssp1 (32%) and CaMKK1 (28%) (Figure 8A and Table 1).

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