Phosphorylation of the S. cerevisiae Cdc25 in response to glucose results in its dissociation from Ras

Gross, Eitan; Goldberg, D.A.; Levitzki, Alexander

doi:10.1038/360762a0

Cited by 84 publications

(53 citation statements)

References 17 publications

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“…It has been reported that the addition of glucose to glucosestarved cells induces the phosphorylation of Cdc25p and changes its localization from membrane to the cytoplasm (Gross et al, 1992a). In order to determine whether Cdc25p is regulated in the same way in response to heat shock, we checked its phosphorylation state and localization.…”

Section: Resultsmentioning

confidence: 99%

Stress induces depletion of Cdc25p and decreases the cAMP producing capability in Saccharomyces cerevisiae

et al. 2004

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In Saccharomyces cerevisiae the cAMP-dependent protein kinase A pathway antagonizes the cellular response to stress. It is shown here that the cellular content of Cdc25p, the upstream activator of Ras and adenylyl cyclase, decays upon various stresses such as heat shock and oxidative and ethanol shocks, whereas its phosphorylation level and its localization are unaffected. In parallel with the reduction of Cdc25p, the maximal capacity of the cell to accumulate cAMP decreases when its feedback regulation is abolished. A deletion of CDC25 prevents this decrease. Paradoxically, in wild-type cells, with normal feedback regulation, the level of cAMP, which is much lower, is not reduced but is rather increased upon stress. These observations are consistent with a role of Cdc25p in sensing and transducing stress to downstream targets, either through a cAMP-independent pathway or by large fluctuations in the cAMP content of the cell.

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Section: Resultsmentioning

confidence: 99%

Stress induces depletion of Cdc25p and decreases the cAMP producing capability in Saccharomyces cerevisiae

et al. 2004

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“…During the course of these studies, we found that a particular inactive variant of Tpk1 exhibited an increased binding to a number of known PKA substrates, including Cdc25 and Cki1 ( Figure 1A). Cdc25 is the guanine nucleotide exchange factor for the Ras proteins in S. cerevisiae, and Cki1 is a choline kinase (Gross et al 1992;Yu et al 2002). In co-immunoprecipitation assays, the Tpk1 KH variant bound significantly more of both of these substrates than did the wild-type protein ( Figure 1, A and B).…”

Section: The Catalytically Inactive Pka Variant Tpk1mentioning

confidence: 99%

“…Most of the targets used above are likely phosphorylated by PKA at more than one position (Gross et al 1992;Reinders et al 1998;Yu et al 2002). For these experiments, we used a fragment of Yak1 that encompassed codons 243-361 and was phosphorylated by PKA at the serine residue at position 295 ( Figure 4C).…”

Section: Both Alterations In Tpk1mentioning

confidence: 99%

Using Substrate-Binding Variants of the cAMP-Dependent Protein Kinase to Identify Novel Targets and a Kinase Domain Important for Substrate Interactions in Saccharomyces cerevisiae

et al. 2006

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Protein kinases mediate much of the signal transduction in eukaryotic cells and defects in kinase function are associated with a variety of human diseases. To understand and correct these defects, we will need to identify the physiologically relevant substrates of these enzymes. The work presented here describes a novel approach to this identification process for the cAMP-dependent protein kinase (PKA) in Saccharomyces cerevisiae. This approach takes advantage of two catalytically inactive PKA variants, Tpk1 K336A/H338A and Tpk1 R324A, that exhibit a stable binding to their substrates. Most protein kinases, including the wildtype PKA, associate with substrates with a relatively low affinity. The binding observed here was specific to substrates and was dependent upon PKA residues known to be important for interactions with peptide substrates. The general utility of this approach was demonstrated by the ability to identify both previously described and novel PKA substrates in S. cerevisiae. Interestingly, the positions of the residues altered in these variants implicated a particular region within the PKA kinase domain, corresponding to subdomain XI, in the binding and/or release of protein substrates. Moreover, the high conservation of the residues altered and, in particular, the invariant nature of the R324 position suggest that this approach might be generally applicable to other protein kinases.

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“…Consistent with these observations, the proline-rich carboxy-terminal region of SOS contains a number of MAPK consensus sites, some of which have been shown to be phosphorylated by this kinase both in vitro and in vivo (Cherniack et al, 1994;Corbalan-Garcia et al, 1996b;Rozakis-Adcock et al, 1995). By analogy to S. cerivisiae where Cdc25 phosphorylation is involved in down regulation of the Ras pathway (Gross et al, 1992), MAP kinase phosphorylation of SOS in mammalian cells is thought to play a similar role. However, while no change in SOS exchange activity has yet been demonstrated in response to MAP kinase phosphorylation, the binding to Grb2 and Shc has been shown to be altered.…”

Section: Introductionmentioning

confidence: 99%

EGF induced SOS phosphorylation in PC12 cells involves P90 RSK-2

1997

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SOS, the guanine nucleotide exchange factor for Ras, becomes phosphorylated on serine and threonine residues following stimulation of cells with growth factors. These phosphorylations may play a role in negative feedback of Ras stimulation and have been shown to be mediated in part by the MAP kinases Erk-1 and Erk-2. Here we show that in addition to MAP kinase, a major mitogen activated kinase for SOS is p90 Rsk-2, a downstream target of MAP kinase. p90 Rsk-2 phosphorylates SOS in an in gel assay and also in solution in vitro. The ability of p90 Rsk-2 to phosphorylate SOS increases greatly following EGF treatment of PC12 cells and is blocked by expression of N17 Ras or treatment with the MEK inhibitor PD98059. Phosphopeptide mapping revealed that the sites phosphorylated by p90 Rsk-2 in vitro were also phosphorylated in intact cells in response to EGF treatment. Several major sites of in vivo phosphorylation correlated with p90 Rsk-2 phosphorylation sites rather than MAP kinase sites. It is therefore likely that p90 Rsk-2 plays an important role in the down regulation of the Ras activation pathway through SOS.

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Phosphorylation of the S. cerevisiae Cdc25 in response to glucose results in its dissociation from Ras

Cited by 84 publications

References 17 publications

Stress induces depletion of Cdc25p and decreases the cAMP producing capability in Saccharomyces cerevisiae

Stress induces depletion of Cdc25p and decreases the cAMP producing capability in Saccharomyces cerevisiae

Using Substrate-Binding Variants of the cAMP-Dependent Protein Kinase to Identify Novel Targets and a Kinase Domain Important for Substrate Interactions in Saccharomyces cerevisiae

EGF induced SOS phosphorylation in PC12 cells involves P90 RSK-2

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