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Protein kinase C controls activation of the DNA integrity checkpoint.

Soriano-Carot M, Quilis I, Bañó MC, Igual JC - Nucleic Acids Res. (2014)

Bottom Line: This modification is a phosphorylation event mediated by Tel1.The expression of different mammalian PKC isoforms at the endogenous level in yeast pkc1 mutant cells revealed that PKCδ is able to activate the DNA integrity checkpoint.Our results indicate that the control of the DNA integrity checkpoint by PKC is a mechanism conserved from yeast to humans.

View Article: PubMed Central - PubMed

Affiliation: Departament de Bioquímica i Biologia Molecular. Universitat de València, 46100 Burjassot (Valencia), Spain.

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Analysis of Rad53 checkpoint kinase activation by a kinase-dead Pkc1 mutant protein. Exponentially growing cultures of the pkc1ts (JC6-3a) strain transformed with a centromeric plasmid containing the PKC1, the PKC1K853R (kinase-dead mutation) gene or an empty vector were split and incubated for 3 h at 25º or 37º followed by 1 h incubation in the absence or presence of 0.2 M HU or 0.04% MMS. Activation of the checkpoint kinase Rad53 was determined by western analysis. The ponceau staining of the membrane is shown as loading control.
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Figure 3: Analysis of Rad53 checkpoint kinase activation by a kinase-dead Pkc1 mutant protein. Exponentially growing cultures of the pkc1ts (JC6-3a) strain transformed with a centromeric plasmid containing the PKC1, the PKC1K853R (kinase-dead mutation) gene or an empty vector were split and incubated for 3 h at 25º or 37º followed by 1 h incubation in the absence or presence of 0.2 M HU or 0.04% MMS. Activation of the checkpoint kinase Rad53 was determined by western analysis. The ponceau staining of the membrane is shown as loading control.

Mentions: Next, we tested whether Pkc1 kinase activity is required for proper checkpoint activation in response to DNA damage. Rad53 activation was checked in pkc1 mutant cells expressing a kinase-dead mutant protein, Pkc1K853R (49). Whereas the expression of the wild-type PKC1 gene was able to sustain Rad53 activation, no Rad53 phosphorylation was observed in the cells expressing the Pkc1K853R mutant protein (Figure 3). This indicates that the kinase activity of Pkc1 is required for checkpoint control in response to DNA damage, suggesting that Pkc1 may act by phosphorylating essential targets for Rad53 activation.


Protein kinase C controls activation of the DNA integrity checkpoint.

Soriano-Carot M, Quilis I, Bañó MC, Igual JC - Nucleic Acids Res. (2014)

Analysis of Rad53 checkpoint kinase activation by a kinase-dead Pkc1 mutant protein. Exponentially growing cultures of the pkc1ts (JC6-3a) strain transformed with a centromeric plasmid containing the PKC1, the PKC1K853R (kinase-dead mutation) gene or an empty vector were split and incubated for 3 h at 25º or 37º followed by 1 h incubation in the absence or presence of 0.2 M HU or 0.04% MMS. Activation of the checkpoint kinase Rad53 was determined by western analysis. The ponceau staining of the membrane is shown as loading control.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4066786&req=5

Figure 3: Analysis of Rad53 checkpoint kinase activation by a kinase-dead Pkc1 mutant protein. Exponentially growing cultures of the pkc1ts (JC6-3a) strain transformed with a centromeric plasmid containing the PKC1, the PKC1K853R (kinase-dead mutation) gene or an empty vector were split and incubated for 3 h at 25º or 37º followed by 1 h incubation in the absence or presence of 0.2 M HU or 0.04% MMS. Activation of the checkpoint kinase Rad53 was determined by western analysis. The ponceau staining of the membrane is shown as loading control.
Mentions: Next, we tested whether Pkc1 kinase activity is required for proper checkpoint activation in response to DNA damage. Rad53 activation was checked in pkc1 mutant cells expressing a kinase-dead mutant protein, Pkc1K853R (49). Whereas the expression of the wild-type PKC1 gene was able to sustain Rad53 activation, no Rad53 phosphorylation was observed in the cells expressing the Pkc1K853R mutant protein (Figure 3). This indicates that the kinase activity of Pkc1 is required for checkpoint control in response to DNA damage, suggesting that Pkc1 may act by phosphorylating essential targets for Rad53 activation.

Bottom Line: This modification is a phosphorylation event mediated by Tel1.The expression of different mammalian PKC isoforms at the endogenous level in yeast pkc1 mutant cells revealed that PKCδ is able to activate the DNA integrity checkpoint.Our results indicate that the control of the DNA integrity checkpoint by PKC is a mechanism conserved from yeast to humans.

View Article: PubMed Central - PubMed

Affiliation: Departament de Bioquímica i Biologia Molecular. Universitat de València, 46100 Burjassot (Valencia), Spain.

Show MeSH
Related in: MedlinePlus