Phosphoproteome dynamics reveal novel ERK1/2 MAP kinase substrates with broad spectrum of functions

Courcelles, Mathieu; Frémin, Christophe; Voisin, Laure; Lemieux, Sébastien; Meloche, Sylvain; Thibault, Pierre

doi:10.1038/msb.2013.25

Cited by 153 publications

(72 citation statements)

References 45 publications

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“…5B). Together with the previous in vivo study that spliceosome phosphorylation is ERK1 dependent (44), the enrichment of spliceosomal proteins in our result further suggested that ERK1 probably contributes to the splicing regulation directly through phosphorylation. In addition, the direct ERK1 substrates could be grouped into several important biological processes: gene expression, metabolism, cellular assembly, cell cycle, transport, and signal transduction (Fig.…”

Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalsupporting

confidence: 89%

“…Third, a full spectrum of kinase-dependent phosphorylation change in vivo is essential. In this study, Courcelles and coworkers have reported the most comprehensive quantitative phosphoproteomics on ERK1 to our knowledge (44), which led to the most overlapping substrates (34 proteins) with our in vitro kinase reactions. It also highlights the importance of highly specific and efficient kinase inhibition in vivo.…”

Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalmentioning

confidence: 85%

“…In the same year, Hattori's group identified 38 proteins using 2D-DIGE, IMAC, and phosphomotif antibodies (43). Recently Thibault and colleagues further expanded the ERK1-dependent repertoire, which contains 155 proteins with 232 phosphorylation sites using a rat cell line (44). In addition, two previously mentioned studies identified in vivo substrates of ERK2 (40) and p38 (34), which also belong to mitogen activation protein kinase (MAPK) family.…”

Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalmentioning

confidence: 99%

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Identification of Extracellular Signal-regulated Kinase 1 (ERK1) Direct Substrates using Stable Isotope Labeled Kinase Assay-Linked Phosphoproteomics

Xue¹,

Wang²,

Cao³

et al. 2014

Molecular & Cellular Proteomics

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Kinase mediated phosphorylation signaling is extensively involved in cellular functions and human diseases, and unraveling phosphorylation networks requires the identification of substrates targeted by kinases, which has remained challenging. We report here a novel proteomic strategy to identify the specificity and direct substrates of kinases by coupling phosphoproteomics with a sensitive stable isotope labeled kinase reaction. A whole cell extract was moderately dephosphorylated and subjected to in vitro kinase reaction under the condition in which 18 O-ATP is the phosphate donor. The phosphorylated proteins are then isolated and identified by mass spectrometry, in which the heavy phosphate (؉85.979 Da) labeled phosphopeptides reveal the kinase specificity. The in vitro phosphorylated proteins with heavy phosphates are further overlapped with in vivo kinase-dependent phosphoproteins for the identification of direct substrates with high confidence. The strategy allowed us to identify 46 phosphorylation sites on 38 direct substrates of extracellular signal-regulated kinase 1, including multiple known substrates and novel substrates, highlighting the ability of this high throughput method for direct kinase substrate screening. Molecular & Cellular

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Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalsupporting

confidence: 89%

Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalmentioning

confidence: 85%

Section: Identifying Direct Substrate Of Erk1 Under the Physiologicalmentioning

confidence: 99%

See 1 more Smart Citation

Identification of Extracellular Signal-regulated Kinase 1 (ERK1) Direct Substrates using Stable Isotope Labeled Kinase Assay-Linked Phosphoproteomics

Xue¹,

Wang²,

Cao³

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Furthermore, many biological processes enriched in both cell and xenograft phosphoproteomes (supplemental Fig. S3B), including protein folding and the cell cycle, are known to be regulated by MAPK/ERK signaling (62,69,70), highlighting the connection between citreoviridin treatment and MAPK/ERK cascade in cancer cells.…”

Section: Discussionmentioning

confidence: 99%

Temporal Phosphoproteome Dynamics Induced by an ATP Synthase Inhibitor Citreoviridin*

Hsu

Wang³

et al. 2015

Molecular & Cellular Proteomics

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Citreoviridin, one of toxic mycotoxins derived from fungal species, can suppress lung cancer cell growth by inhibiting the activity of ectopic ATP synthase, but has limited effect on normal cells. However, the mechanism of citreoviridin triggering dynamic molecular responses in cancer cells remains unclear. Here, we performed temporal phosphoproteomics to elucidate the dynamic changes after citreoviridin treatment in cells and xenograft model. We identified a total of 829 phosphoproteins and demonstrated that citreoviridin treatment affects protein folding, cell cycle, and cytoskeleton function. Furthermore, response network constructed by mathematical modeling shows the relationship between the phosphorylated heat shock protein 90 β and mitogen-activated protein kinase signaling pathway. This work describes that citreoviridin suppresses cancer cell growth and mitogenactivated protein kinase/extracellular signal-regulated kinase signaling by site-specific dephosphorylation of HSP90AB1 on Serine 255 and provides perspectives in cancer therapeutic strategies. Molecular & Cellular

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“…Ras first activates Polehole, which in turn phosphorylates Dsor1 to activate it. Dsor1 can then dually phosphorylate and activate Rl, which in turn can then phosphorylate and modulate a wide range of substrates throughout the cell, including transcription factors to affect target gene expression (Courcelles et al, 2013). Several proteins from the EGFR-Ras-MAPK pathway have been identified to directly interact with Wnt signaling components, in particular ERK family proteins (Červenka et al, 2011;Hoschuetzky et al, 1994;Krejci et al, 2012;Ding et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Ras-activated Dsor1 promotes Wnt signaling in Drosophila development

Hall

Verheyen

2015

Journal of Cell Science

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Wnt/Wingless (Wg) and Ras-MAPK signaling both play fundamental roles in growth and cell fate determination, and when dysregulated, can lead to tumorigenesis. Several conflicting modes of interaction between Ras-MAPK and Wnt signaling have been identified in specific cellular contexts, causing synergistic or antagonistic effects on target genes. We find novel evidence that the Drosophila homolog of the dual specificity kinases MEK1/2 (also known as MAP2K1/2), Downstream of Raf1 (Dsor1), is required for Wnt signaling. Knockdown of Dsor1 results in loss of Wg target gene expression, as well as reductions in stabilized Armadillo (Arm; Drosophila β-catenin). We identify a close physical interaction between Dsor1 and Arm, and find that catalytically inactive Dsor1 causes a reduction in active Arm. These results suggest that Dsor1 normally counteracts the Axin-mediated destruction of Arm. We find that Ras-Dsor1 activity is independent of upstream activation by EGFR, and instead it appears to be activated by the insulin-like growth factor receptor to promote Wg signaling. Taken together, our results suggest that there is a new crosstalk pathway between insulin and Wg signaling that is mediated by Dsor1.

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Phosphoproteome dynamics reveal novel ERK1/2 MAP kinase substrates with broad spectrum of functions

Abstract: Quantitative phosphoproteomics was used to measure the dynamic changes in phosphorylation of ERK1/2 MAP kinases consensus sequences in epithelial cells and to identify 128 novel candidate substrates involved in a broad spectrum of biological functions.

Cited by 153 publications

References 45 publications

Identification of Extracellular Signal-regulated Kinase 1 (ERK1) Direct Substrates using Stable Isotope Labeled Kinase Assay-Linked Phosphoproteomics

Identification of Extracellular Signal-regulated Kinase 1 (ERK1) Direct Substrates using Stable Isotope Labeled Kinase Assay-Linked Phosphoproteomics

Temporal Phosphoproteome Dynamics Induced by an ATP Synthase Inhibitor Citreoviridin*

Ras-activated Dsor1 promotes Wnt signaling in Drosophila development

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