Mechanisms of site‐specific dephosphorylation and kinase opposition imposed by PP2A regulatory subunits
Open accessAbstract
PP2A is an essential protein phosphatase that regulates most cellular processes through the formation of holoenzymes containing distinct regulatory B‐subunits. Only a limited number of PP2A‐regulated phosphorylation sites are known. This hampers our understanding of the mechanisms of site‐specific dephosphorylation and of its tumor suppressor functions. Here, we develop phosphoproteomic strategies for global substrate identification of PP2A‐B56 and PP2A‐B55 holoenzymes. Strikingly, we find that B‐subunits directly affect the dephosphorylation site preference of the PP2A catalytic subunit, resulting in unique patterns of kinase opposition. For PP2A‐B56, these patterns are further modulated by affinity and position of B56 binding motifs. Our screens identify phosphorylation sites in the cancer target ADAM17 that are regulated through a conserved B56 binding site. Binding of PP2A‐B56 to ADAM17 protease decreases growth factor signaling and tumor development in mice. This work provides a roadmap for the identification of phosphatase substrates and reveals unexpected mechanisms governing PP2A dephosphorylation site specificity and tumor suppressor function.
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B56α interacts with up to 100 cellular target proteins.
“B56α interacts with up to 100 cellular target proteins (Supplementary Data 1 and 10,11) among which, it is best known by its capacity to inhibit oncogenic activity of MYC12,13.”
From Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A (2023) · cites this paper as [11]
The Results section, “Development of a specific PP2A-B56 inhibitor” (page unavailable), reports cellular proteins that interact with YFP-B56α, supporting the interaction part of the claim. It does not give a count approaching 100 B56α interactors. The 651 proteins reported in “Identification of PP2A-dependent phosphorylation sites” are proteins with increased phosphorylation, not a count of B56α-binding targets. (LLM, conf=0.85)
Experimental — accuracy not yet measuredB′ subunits require an intact A subunit for binding, as deduced from in vitro experiments and mouse models.
“Limited insights have been gained from the B’ subunits, which require an intact A subunit for binding, as deduced from in vitro experiments and mouse models.”
From Targeting PP2A in cancer: an underrated option (2025) · cites this paper as [59]
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Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A
Pavic, K., Gupta, N., Omella, J. D., Derua, R., Aakula, A., Huhtaniemi, R., Määttä, J. A., Höfflin, N., Okkeri, J., Wang, Z., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-36693-9 · Cited by 36 · see citing works
“The data show that the CIP2A(1-560) residues cross-linked to B56α are located primarily in the N-terminal head domain (K18, K21, E23, E34, K40) with an additional residue at the C-terminus (K490) (Fig.”
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Targeting PP2A in cancer: an underrated option
Sarais, F., Krempien, F., Koehn, C., Brewing, L., Classen, C. F., Walter, M., & Shokraie, O. (2025) · Journal of Experimental & Clinical Cancer Research · doi:10.1186/s13046-025-03560-y · Cited by 8 · see citing works
- Authors
- Kruse, T., Gnosa, S. P., Nasa, I., Garvanska, D. H., Hein, J. B., Nguyen, H., Samsøe‐Petersen, J., Lopez‐Mendez, B., Hertz, E. P. T., Schwarz, J., Pena, H. S., Nikodemus, D., Kveiborg, M., Kettenbach, A. N., & Nilsson, J.
- Affiliations
- University of Copenhagen (DK)
- Dartmouth College (US)
- Corresponding author
- Kveiborg, Marie
- Kettenbach, Arminja N
- Nilsson, Jakob
- ORCID
- Venue
- The EMBO Journal
- Published
- 2020-05-13
- Volume
- 39
- Issue
- 13
- Pages
- e103695-e103695
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC7327492
- OpenAlex
- W3024517740
- Cited by
- 97
- References
- 58
- Citations by year
- 2026: 10 · 2025: 12 · 2024: 16 · 2023: 25 · 2022: 7 · 2021: 17 · 2020: 10
- FWCI
- 4.78
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- The EMBO Journal (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
- Funders
- Novo Nordisk Foundation (NNF14CC0001, NNF18OC0053124)
- Kræftens Bekæmpelse (R146‐A9211‐16‐S2, R167‐A10951‐17‐S2)
- Independent Research Fund Denmark (8021‐00101B, 7016 00086)
- NIH/NIGMS (R35GM119455, P20GM113132)
- NIH (S10‐OD016212)
- Marie Sklodowska Curie Grant (798716)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
- Keywords
- protein phosphatase 2A, PP2A-B56, PP2A-B55, phosphoproteomics, ADAM17, tumor suppressor function, growth factor signaling, carcinogenesis
- MeSH terms
- ADAM17 Protein, Animals, Binding Sites, HeLa Cells, Humans, Phosphorylation, Amino Acid Motifs, Mice, Protein Phosphatase 2
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