CoChat
Sign Up
Cited by 8514-3-3 protein interactionsProtein Kinase Regulation and GTPase SignalingProtein Tyrosine Phosphatases

Phosphoproteomic analysis identifies the tumor suppressor PDCD4 as a RSK substrate negatively regulated by 14-3-3

Open access
Galan, Jacob A., Geraghty, Kathryn M., Lavoie, Geneviève, Kanshin, Evgeny, Tcherkezian, Joseph, Calabrese, Viviane, Jeschke, Grace R., Turk, Benjamin E. +4 more·2014·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.1405601111PMC PMC4115529
Open this paper in CoChat — free account

Abstract

The Ras/MAPK signaling cascade regulates various biological functions, including cell growth and proliferation. As such, this pathway is frequently deregulated in several types of cancer, including most cases of melanoma. RSK (p90 ribosomal S6 kinase) is a MAPK-activated protein kinase required for melanoma growth and proliferation, but relatively little is known about its exact function and the nature of its substrates. Herein, we used a quantitative phosphoproteomics approach to define the signaling networks regulated by RSK in melanoma. To more accurately predict direct phosphorylation substrates, we defined the RSK consensus phosphorylation motif and found significant overlap with the binding consensus of 14-3-3 proteins. We thus characterized the phospho-dependent 14-3-3 interactome in melanoma cells and found that a large proportion of 14-3-3 binding proteins are also potential RSK substrates. Our results show that RSK phosphorylates the tumor suppressor PDCD4 (programmed cell death protein 4) on two serine residues (Ser76 and Ser457) that regulate its subcellular localization and interaction with 14-3-3 proteins. We found that 14-3-3 binding promotes PDCD4 degradation, suggesting an important role for RSK in the inactivation of PDCD4 in melanoma. In addition to this tumor suppressor, our results suggest the involvement of RSK in a vast array of unexplored biological functions with relevance in oncogenesis.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1073/pnas.1405601111

Cites

6 of 6 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • 14-3-3 protein interacted with PDCD4 protein phosphorylated at S67, S76, and/or S457, sequestering PDCD4 into the cytoplasm and stimulating degradation of the protein in the proteasome system.

    “They showed that 14-3-3 protein was co-immuno-precipitated with PDCD4-protein when RSK was activated by the PMA treatment of melanoma cells and conclude that 14-3-3 protein interacted with PDCD4 protein phosphorylated at S67, S76, and/or S457 sequestering the PDCD4-protein into the cytoplasm and stimulated the degradation of the protein in the proteasome system [71].”

    From Control Mechanisms of the Tumor Suppressor PDCD4: Expression and Functions (2019) · cites this paper as [71]

    The Results say that Ser76 and Ser457, but not Ser67, are required for regulated 14-3-3 binding. They also report nuclear accumulation of PDCD4 after RSK activation, not sequestration in the cytoplasm. The paper does support the claim that 14-3-3 binding promotes PDCD4 degradation. (LLM, conf=0.90)

    Checked by a reasoning model — may be wrong.

    Experimental — accuracy not yet measured

  • The consensus sequence of RSK is RXRXXS/T, the same as that of p70S6K1, and RSK phosphorylates Ser76 and Ser457, as well as Ser67, of human PDCD4 protein in melanoma cells.

    “It was shown that the consensus sequence of p90 ribosomal S6 kinase (RSK) is RXRXXS/T as same as that of p70S6K1 and the enzyme phosphorylates, the Ser76 and Ser457, as well as Ser67 of human PDCD4-protein in melanoma cells [71].”

    From Control Mechanisms of the Tumor Suppressor PDCD4: Expression and Functions (2019) · cites this paper as [71]

    The Discussion reports that RSK phosphorylates PDCD4 at Ser76 and Ser457, including Ser457 in melanoma cells. The Results describe Ser67 only as a low-confidence potential 14-3-3 binding site, not as a site phosphorylated by RSK. The paper also describes a more specific RSK motif but does not establish that it is identical to the p70S6K1 motif. (LLM, conf=0.91)

    Experimental — accuracy not yet measured

  • RSK is activated via the mitogen-induced MAPK pathway.

    “RSK is activated via mitogen-induced mitogen activated protein kinase (MAPK) pathway [71].”

    From Control Mechanisms of the Tumor Suppressor PDCD4: Expression and Functions (2019) · cites this paper as [71]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 26

  1. [3]Michaloglou C, Vredeveld LC, Mooi WJ, Peeper DS. BRAF(E600) in benign and malignant human tumours. Oncogene. 2008;27(7):877–895. doi: 10.1038/sj.onc.1210704. [DOI] [PubMed] [Google Scholar] · DOI
  2. [4]Fecher LA, Amaravadi RK, Flaherty KT. The MAPK pathway in melanoma. Curr Opin Oncol. 2008;20(2):183–189. doi: 10.1097/CCO.0b013e3282f5271c. [DOI] [PubMed] [Google Scholar] · DOI
  3. [5]Cargnello M, Roux PP. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev. 2011;75(1):50–83. doi: 10.1128/MMBR.00031-10. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI · Paper page
  4. [6]Romeo Y, Zhang X, Roux PP. Regulation and function of the RSK family of protein kinases. Biochem J. 2012;441(2):553–569. doi: 10.1042/BJ20110289. [DOI] [PubMed] [Google Scholar] · DOI
  5. [7]Smith JA, et al. Identification of the first specific inhibitor of p90 ribosomal S6 kinase (RSK) reveals an unexpected role for RSK in cancer cell proliferation. Cancer Res. 2005;65(3):1027–1034. [PubMed] [Google Scholar]
  6. [8]Clark DE, et al. The serine/threonine protein kinase, p90 ribosomal S6 kinase, is an important regulator of prostate cancer cell proliferation. Cancer Res. 2005;65(8):3108–3116. doi: 10.1158/0008-5472.CAN-04-3151. [DOI] [PubMed] [Google Scholar] · DOI
  7. [9]Old WM, et al. Functional proteomics identifies targets of phosphorylation by B-Raf signaling in melanoma. Mol Cell. 2009;34(1):115–131. doi: 10.1016/j.molcel.2009.03.007. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  8. [10]Romeo Y, et al. RSK regulates activated BRAF signalling to mTORC1 and promotes melanoma growth. Oncogene. 2013;32(24):2917–2926. doi: 10.1038/onc.2012.312. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  9. [11]Zhao J, Meyerkord CL, Du Y, Khuri FR, Fu H. 14-3-3 proteins as potential therapeutic targets. Semin Cell Dev Biol. 2011;22(7):705–712. doi: 10.1016/j.semcdb.2011.09.012. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  10. [12]Gardino AK, Yaffe MB. 14-3-3 proteins as signaling integration points for cell cycle control and apoptosis. Semin Cell Dev Biol. 2011;22(7):688–695. doi: 10.1016/j.semcdb.2011.09.008. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  11. [13]Moritz A, et al. Akt-RSK-S6 kinase signaling networks activated by oncogenic receptor tyrosine kinases. Sci Signal. 2010;3(136):ra64. doi: 10.1126/scisignal.2000998. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  12. [14]Leighton IA, Dalby KN, Caudwell FB, Cohen PT, Cohen P. Comparison of the specificities of p70 S6 kinase and MAPKAP kinase-1 identifies a relatively specific substrate for p70 S6 kinase: The N-terminal kinase domain of MAPKAP kinase-1 is essential for peptide phosphorylation. FEBS Lett. 1995;375(3):289–293. doi: 10.1016/0014-5793(95)01170-j. [DOI] [PubMed] [Google Scholar] · DOI
  13. [15]Chen C, Turk BE. Analysis of serine-threonine kinase specificity using arrayed positional scanning peptide libraries. Curr Protoc Mol Biol. 2010;Chap 18:Unit 18.14. doi: 10.1002/0471142727.mb1814s91. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  14. [16]Obenauer JC, Cantley LC, Yaffe MB. Scansite 2.0: Proteome-wide prediction of cell signaling interactions using short sequence motifs. Nucleic Acids Res. 2003;31(13):3635–3641. doi: 10.1093/nar/gkg584. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  15. [17]Dorrello NV, et al. S6K1- and betaTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. Science. 2006;314(5798):467–471. doi: 10.1126/science.1130276. [DOI] [PubMed] [Google Scholar] · DOI · Paper page
  16. [19]Brunet A, et al. 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. J Cell Biol. 2002;156(5):817–828. doi: 10.1083/jcb.200112059. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  17. [20]Masters SC, Fu H. 14-3-3 proteins mediate an essential anti-apoptotic signal. J Biol Chem. 2001;276(48):45193–45200. doi: 10.1074/jbc.M105971200. [DOI] [PubMed] [Google Scholar] · DOI
  18. [21]Saha M, et al. RSK phosphorylates SOS1 creating 14-3-3-docking sites and negatively regulating MAPK activation. Biochem J. 2012;447(1):159–166. doi: 10.1042/BJ20120938. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  19. [22]Lankat-Buttgereit B, Göke R. The tumour suppressor Pdcd4: Recent advances in the elucidation of function and regulation. Biol Cell. 2009;101(6):309–317. doi: 10.1042/BC20080191. [DOI] [PubMed] [Google Scholar] · DOI
  20. [24]Roux PP, et al. RAS/ERK signaling promotes site-specific ribosomal protein S6 phosphorylation via RSK and stimulates cap-dependent translation. J Biol Chem. 2007;282(19):14056–14064. doi: 10.1074/jbc.M700906200. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  21. [26]Blees JS, et al. Development of a high-throughput cell-based reporter assay to identify stabilizers of tumor suppressor Pdcd4. J Biomol Screen. 2010;15(1):21–29. doi: 10.1177/1087057109351028. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  22. [27]Romeo Y, Roux PP. Paving the way for targeting RSK in cancer. Expert Opin Ther Targets. 2011;15(1):5–9. doi: 10.1517/14728222.2010.531014. [DOI] [PubMed] [Google Scholar] · DOI
  23. [29]Ballif BA, Cao Z, Schwartz D, Carraway KL, 3rd, Gygi SP. Identification of 14-3-3epsilon substrates from embryonic murine brain. J Proteome Res. 2006;5(9):2372–2379. doi: 10.1021/pr060206k. [DOI] [PubMed] [Google Scholar] · DOI
  24. [30]Yaffe MB, et al. The structural basis for 14-3-3:phosphopeptide binding specificity. Cell. 1997;91(7):961–971. doi: 10.1016/s0092-8674(00)80487-0. [DOI] [PubMed] [Google Scholar] · DOI
  25. [31]Mok J, et al. Deciphering protein kinase specificity through large-scale analysis of yeast phosphorylation site motifs. Sci Signal. 2010;3(109):ra12. doi: 10.1126/scisignal.2000482. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI
  26. [32]Roux PP, Ballif BA, Anjum R, Gygi SP, Blenis J. Tumor-promoting phorbol esters and activated Ras inactivate the tuberous sclerosis tumor suppressor complex via p90 ribosomal S6 kinase. Proc Natl Acad Sci USA. 2004;101(37):13489–13494. doi: 10.1073/pnas.0405659101. [DOI] [PMC free article] [PubMed] [Google Scholar] · DOI

Cited by

Sorted by citations

Authors
Galan, J. A., Geraghty, K. M., Lavoie, G., Kanshin, E., Tcherkezian, J., Calabrese, V., Jeschke, G. R., Turk, B. E., Ballif, B. A., Blenis, J., Thibault, P., & Roux, P. P.
Affiliations
  • Institute for Research in Immunology and Cancer (CA)
  • Harvard University (US)
  • Yale University (US)
  • University of Vermont (US)
ORCID
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2014-07-07
Volume
111
Issue
29
Pages
E2918-27
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.1405601111
PMC
PMC4115529
OpenAlex
W2045424472
Cited by
85
References
6
Citations by year
2026: 3 · 2025: 4 · 2024: 11 · 2023: 3 · 2022: 6 · 2021: 4 · 2020: 12 · 2019: 12 · 2018: 8 · 2017: 8 · 2016: 7 · 2015: 4 · 2014: 3
FWCI
3.01
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Canadian Institutes of Health Research
  • National Cancer Institute (R01CA046595)
  • National Institute of General Medical Sciences (R01GM104047, P20GM103449)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › 14-3-3 protein interactions
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
Keywords
PDCD4, RSK, 14-3-3 proteins, melanoma, RAS/MAPK pathway, protein phosphorylation, protein degradation, tumor suppressor inactivation, protein-protein interaction, subcellular localization
MeSH terms
Amino Acid Sequence, Cell Line, Cell Nucleus, Humans, Melanoma, Models, Biological, Molecular Sequence Data, Phosphoproteins, Phosphorylation, Phosphoserine, Protein Binding, Substrate Specificity, Consensus Sequence, RNA-Binding Proteins, Peptide Library, Proteome, Amino Acid Motifs, Protein Transport, Tumor Suppressor Proteins, Ribosomal Protein S6 Kinases, 90-kDa, Proteomics, 14-3-3 Proteins, Apoptosis Regulatory Proteins, Proteolysis

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review