Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A

Karolina Pavic; Nikhil Gupta; Judit Domènech Omella; Rita Derua; Anna Aakula; Riikka Huhtaniemi; Juha A. Määttä; Nico Höfflin; Juha Okkeri; Zhizhi Wang; Otto Kauko; Roosa Varjus; Henrik Honkanen; Daniel Abankwa; Maja Köhn; Vesa P. Hytönen; Wenqing Xu; Jakob Nilsson; Rebecca Page; Veerle Janssens; Alexander Leitner; Jukka Westermarck

doi:10.1038/s41467-023-36693-9

Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A

Karolina Pavic, Nikhil Gupta, Judit Domènech Omella, Rita Derua, Anna Aakula, Riikka Huhtaniemi, Juha A. Määttä, Nico Höfflin, Juha Okkeri, Zhizhi Wang, Otto Kauko, Roosa Varjus, Henrik Honkanen, Daniel Abankwa, Maja Köhn, Vesa P. Hytönen, Wenqing Xu, Jakob Nilsson, Rebecca Page, Veerle JanssensAlexander Leitner, Jukka Westermarck

Department of Clinical Chemistry

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Abstract

The protein phosphatase 2A (PP2A) heterotrimer PP2A-B56α is a human tumour suppressor. However, the molecular mechanisms inhibiting PP2A-B56α in cancer are poorly understood. Here, we report molecular level details and structural mechanisms of PP2A-B56α inhibition by an oncoprotein CIP2A. Upon direct binding to PP2A-B56α trimer, CIP2A displaces the PP2A-A subunit and thereby hijacks both the B56α, and the catalytic PP2Ac subunit to form a CIP2A-B56α-PP2Ac pseudotrimer. Further, CIP2A competes with B56α substrate binding by blocking the LxxIxE-motif substrate binding pocket on B56α. Relevant to oncogenic activity of CIP2A across human cancers, the N-terminal head domain-mediated interaction with B56α stabilizes CIP2A protein. Functionally, CRISPR/Cas9-mediated single amino acid mutagenesis of the head domain blunted MYC expression and MEK phosphorylation, and abrogated triple-negative breast cancer in vivo tumour growth. Collectively, we discover a unique multi-step hijack and mute protein complex regulation mechanism resulting in tumour suppressor PP2A-B56α inhibition. Further, the results unfold a structural determinant for the oncogenic activity of CIP2A, potentially facilitating therapeutic modulation of CIP2A in cancer and other diseases.

Original language	English
Article number	1143
Journal	Nature Communications
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41467-023-36693-9
Publication status	Published - 2023
Publication type	A1 Journal article-refereed

Funding

Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP). Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP).

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Publication forum level 3

ASJC Scopus subject areas

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General
General Physics and Astronomy

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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Pavic, K., Gupta, N., Omella, J. D., Derua, R., Aakula, A., Huhtaniemi, R., Määttä, J. A., Höfflin, N., Okkeri, J., Wang, Z., Kauko, O., Varjus, R., Honkanen, H., Abankwa, D., Köhn, M., Hytönen, V. P., Xu, W., Nilsson, J., Page, R., ... Westermarck, J. (2023). Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A. Nature Communications, 14(1), Article 1143. https://doi.org/10.1038/s41467-023-36693-9

@article{ca539caf871f4f15b52cfeddc2457228,

title = "Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A",

abstract = "The protein phosphatase 2A (PP2A) heterotrimer PP2A-B56α is a human tumour suppressor. However, the molecular mechanisms inhibiting PP2A-B56α in cancer are poorly understood. Here, we report molecular level details and structural mechanisms of PP2A-B56α inhibition by an oncoprotein CIP2A. Upon direct binding to PP2A-B56α trimer, CIP2A displaces the PP2A-A subunit and thereby hijacks both the B56α, and the catalytic PP2Ac subunit to form a CIP2A-B56α-PP2Ac pseudotrimer. Further, CIP2A competes with B56α substrate binding by blocking the LxxIxE-motif substrate binding pocket on B56α. Relevant to oncogenic activity of CIP2A across human cancers, the N-terminal head domain-mediated interaction with B56α stabilizes CIP2A protein. Functionally, CRISPR/Cas9-mediated single amino acid mutagenesis of the head domain blunted MYC expression and MEK phosphorylation, and abrogated triple-negative breast cancer in vivo tumour growth. Collectively, we discover a unique multi-step hijack and mute protein complex regulation mechanism resulting in tumour suppressor PP2A-B56α inhibition. Further, the results unfold a structural determinant for the oncogenic activity of CIP2A, potentially facilitating therapeutic modulation of CIP2A in cancer and other diseases.",

author = "Karolina Pavic and Nikhil Gupta and Omella, {Judit Dom{\`e}nech} and Rita Derua and Anna Aakula and Riikka Huhtaniemi and M{\"a}{\"a}tt{\"a}, {Juha A.} and Nico H{\"o}fflin and Juha Okkeri and Zhizhi Wang and Otto Kauko and Roosa Varjus and Henrik Honkanen and Daniel Abankwa and Maja K{\"o}hn and Hyt{\"o}nen, {Vesa P.} and Wenqing Xu and Jakob Nilsson and Rebecca Page and Veerle Janssens and Alexander Leitner and Jukka Westermarck",

note = "Funding Information: Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP). Funding Information: Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP). Publisher Copyright: {\textcopyright} 2023, The Author(s).",

year = "2023",

doi = "10.1038/s41467-023-36693-9",

language = "English",

volume = "14",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Research",

number = "1",

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Pavic, K, Gupta, N, Omella, JD, Derua, R, Aakula, A, Huhtaniemi, R, Määttä, JA, Höfflin, N, Okkeri, J, Wang, Z, Kauko, O, Varjus, R, Honkanen, H, Abankwa, D, Köhn, M, Hytönen, VP, Xu, W, Nilsson, J, Page, R, Janssens, V, Leitner, A & Westermarck, J 2023, 'Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A', Nature Communications, vol. 14, no. 1, 1143. https://doi.org/10.1038/s41467-023-36693-9

TY - JOUR

T1 - Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A

AU - Pavic, Karolina

AU - Gupta, Nikhil

AU - Omella, Judit Domènech

AU - Derua, Rita

AU - Aakula, Anna

AU - Huhtaniemi, Riikka

AU - Määttä, Juha A.

AU - Höfflin, Nico

AU - Okkeri, Juha

AU - Wang, Zhizhi

AU - Kauko, Otto

AU - Varjus, Roosa

AU - Honkanen, Henrik

AU - Abankwa, Daniel

AU - Köhn, Maja

AU - Hytönen, Vesa P.

AU - Xu, Wenqing

AU - Nilsson, Jakob

AU - Page, Rebecca

AU - Janssens, Veerle

AU - Leitner, Alexander

AU - Westermarck, Jukka

N1 - Funding Information: Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP). Funding Information: Authors acknowledge the Turku Proteomics Facility, Cell Imaging and Cytometry, and Genome Editing Core at Turku Bioscience Centre, and Turku Centre for Disease Modelling at the University of Turku; all supported by Biocenter Finland. Taina Kalevo-Mattila is acknowledged for excellent technical support, as well as the entire Turku Bioscience personnel for excellent working environment. AL would like to thank Ruedi Aebersold (ETH Zurich) for access to infrastructure and instrumentation. The use of the facilities and expertize of the Protein Service core facility of the Tampere University and a member of Biocenter Finland, is gratefully acknowledged. The authors further want to thank Saverio Minucci for generously providing PLA reagents, Stephen Dilworth for PP2A antibodies, Wolfgang Peti for insightful structural biology advice, and Kari Kurppa for help with Crispr/Cas9 targeting. The work was supported by funding from Sigrid Juselius Foundation (J.W.), Academy of Finland (331237; J.W.)(331946; V.H.), Jane and Aatos Erkko Foundation (J.W.), Finnish Cancer Associations (J.W.), Finnish Cultural Foundation (K.P.), Albin Johanssons Foundation (K.P.), F.W.O. - Vlaanderen (Research Foundation – Flanders) (G0B0116N, G0B1719N, and 1S77521N) (V.J., J.D.O.), KU Leuven (C24/17/073) (V.J., R.D.), Belgian Foundation against Cancer (FA/2020/1330)(V.J.) and NIH NIGMS (R01GM144483)(RP). Publisher Copyright: © 2023, The Author(s).

PY - 2023

Y1 - 2023

N2 - The protein phosphatase 2A (PP2A) heterotrimer PP2A-B56α is a human tumour suppressor. However, the molecular mechanisms inhibiting PP2A-B56α in cancer are poorly understood. Here, we report molecular level details and structural mechanisms of PP2A-B56α inhibition by an oncoprotein CIP2A. Upon direct binding to PP2A-B56α trimer, CIP2A displaces the PP2A-A subunit and thereby hijacks both the B56α, and the catalytic PP2Ac subunit to form a CIP2A-B56α-PP2Ac pseudotrimer. Further, CIP2A competes with B56α substrate binding by blocking the LxxIxE-motif substrate binding pocket on B56α. Relevant to oncogenic activity of CIP2A across human cancers, the N-terminal head domain-mediated interaction with B56α stabilizes CIP2A protein. Functionally, CRISPR/Cas9-mediated single amino acid mutagenesis of the head domain blunted MYC expression and MEK phosphorylation, and abrogated triple-negative breast cancer in vivo tumour growth. Collectively, we discover a unique multi-step hijack and mute protein complex regulation mechanism resulting in tumour suppressor PP2A-B56α inhibition. Further, the results unfold a structural determinant for the oncogenic activity of CIP2A, potentially facilitating therapeutic modulation of CIP2A in cancer and other diseases.

AB - The protein phosphatase 2A (PP2A) heterotrimer PP2A-B56α is a human tumour suppressor. However, the molecular mechanisms inhibiting PP2A-B56α in cancer are poorly understood. Here, we report molecular level details and structural mechanisms of PP2A-B56α inhibition by an oncoprotein CIP2A. Upon direct binding to PP2A-B56α trimer, CIP2A displaces the PP2A-A subunit and thereby hijacks both the B56α, and the catalytic PP2Ac subunit to form a CIP2A-B56α-PP2Ac pseudotrimer. Further, CIP2A competes with B56α substrate binding by blocking the LxxIxE-motif substrate binding pocket on B56α. Relevant to oncogenic activity of CIP2A across human cancers, the N-terminal head domain-mediated interaction with B56α stabilizes CIP2A protein. Functionally, CRISPR/Cas9-mediated single amino acid mutagenesis of the head domain blunted MYC expression and MEK phosphorylation, and abrogated triple-negative breast cancer in vivo tumour growth. Collectively, we discover a unique multi-step hijack and mute protein complex regulation mechanism resulting in tumour suppressor PP2A-B56α inhibition. Further, the results unfold a structural determinant for the oncogenic activity of CIP2A, potentially facilitating therapeutic modulation of CIP2A in cancer and other diseases.

U2 - 10.1038/s41467-023-36693-9

DO - 10.1038/s41467-023-36693-9

M3 - Article

C2 - 36854761

AN - SCOPUS:85149153582

SN - 2041-1723

VL - 14

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 1143

ER -

Structural mechanism for inhibition of PP2A-B56α and oncogenicity by CIP2A

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