Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles

Melissa R. Marzahn; Suresh Marada; Jihun Lee; Amanda Nourse; Sophia Kenrick; Huaying Zhao; Gili Ben-Nissan; Regina Maria Kolaitis; Jennifer L. Peters; Stanley Pounds; Wesley J. Errington; Gilbert G. Privé; J. Paul Taylor; Michal Sharon; Peter Schuck; Stacey K. Ogden; Tanja Mittag

doi:10.15252/embj.201593169

Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles

Melissa R. Marzahn, Suresh Marada, Jihun Lee, Amanda Nourse, Sophia Kenrick, Huaying Zhao, Gili Ben-Nissan, Regina Maria Kolaitis, Jennifer L. Peters, Stanley Pounds, Wesley J. Errington, Gilbert G. Privé, J. Paul Taylor, Michal Sharon, Peter Schuck, Stacey K. Ogden, Tanja Mittag

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

132 Scopus citations

Abstract

Membrane-less organelles in cells are large, dynamic protein/protein or protein/RNA assemblies that have been reported in some cases to have liquid droplet properties. However, the molecular interactions underlying the recruitment of components are not well understood. Herein, we study how the ability to form higher-order assemblies influences the recruitment of the speckle-type POZ protein (SPOP) to nuclear speckles. SPOP, a cullin-3-RING ubiquitin ligase (CRL3) substrate adaptor, self-associates into higher-order oligomers; that is, the number of monomers in an oligomer is broadly distributed and can be large. While wild-type SPOP localizes to liquid nuclear speckles, self-association-deficient SPOP mutants have a diffuse distribution in the nucleus. SPOP oligomerizes through its BTB and BACK domains. We show that BTB-mediated SPOP dimers form linear oligomers via BACK domain dimerization, and we determine the concentration-dependent populations of the resulting oligomeric species. Higher-order oligomerization of SPOP stimulates CRL3^SPOP ubiquitination efficiency for its physiological substrate Gli3, suggesting that nuclear speckles are hotspots of ubiquitination. Dynamic, higher-order protein self-association may be a general mechanism to concentrate functional components in membrane-less cellular bodies.

Original language	English (US)
Pages (from-to)	1254-1275
Number of pages	22
Journal	EMBO Journal
Volume	35
Issue number	12
DOIs	https://doi.org/10.15252/embj.201593169
State	Published - Jun 15 2016

Bibliographical note

Funding Information:
We thank BA Schulman, WK Pierce, EW Martin, and CR Grace for valuable discussions and N Clark for preliminary data collection. We also thank H Haecker (St. Jude Children's Research Hospital). This work was funded by a V Foundation Scholar Grant (T.M.), R01GM112846 (T.M.), R01GM101087 (S.K.O.), the National Cancer Institute Cancer Center Support Grant P30CA21765 (at St. Jude Children's Research Hospital), the American Lebanese Syrian Associated Charities (to J.P.T., S.K.O., and T.M.), and the Intramural Program of the National Institute of Biomedical Imaging and Bioengineering (P.S.). Microscopy images were acquired at the Cell & Tissue Imaging Center at St. Jude Children's Research Hospital.

Publisher Copyright:
© 2016 The Authors. Published under the terms of the CC BY NC ND 4.0 license

Keywords

isodesmic self-association
membrane-less organelle
prostate cancer
speckle-type POZ protein
ubiquitin ligase

UN SDGs

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

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10.15252/embj.201593169

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Marzahn, M. R., Marada, S., Lee, J., Nourse, A., Kenrick, S., Zhao, H., Ben-Nissan, G., Kolaitis, R. M., Peters, J. L., Pounds, S., Errington, W. J., Privé, G. G., Taylor, J. P., Sharon, M., Schuck, P., Ogden, S. K., & Mittag, T. (2016). Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles. EMBO Journal, 35(12), 1254-1275. https://doi.org/10.15252/embj.201593169

Marzahn, MR, Marada, S, Lee, J, Nourse, A, Kenrick, S, Zhao, H, Ben-Nissan, G, Kolaitis, RM, Peters, JL, Pounds, S, Errington, WJ, Privé, GG, Taylor, JP, Sharon, M, Schuck, P, Ogden, SK & Mittag, T 2016, 'Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles', EMBO Journal, vol. 35, no. 12, pp. 1254-1275. https://doi.org/10.15252/embj.201593169

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title = "Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles",

abstract = "Membrane-less organelles in cells are large, dynamic protein/protein or protein/RNA assemblies that have been reported in some cases to have liquid droplet properties. However, the molecular interactions underlying the recruitment of components are not well understood. Herein, we study how the ability to form higher-order assemblies influences the recruitment of the speckle-type POZ protein (SPOP) to nuclear speckles. SPOP, a cullin-3-RING ubiquitin ligase (CRL3) substrate adaptor, self-associates into higher-order oligomers; that is, the number of monomers in an oligomer is broadly distributed and can be large. While wild-type SPOP localizes to liquid nuclear speckles, self-association-deficient SPOP mutants have a diffuse distribution in the nucleus. SPOP oligomerizes through its BTB and BACK domains. We show that BTB-mediated SPOP dimers form linear oligomers via BACK domain dimerization, and we determine the concentration-dependent populations of the resulting oligomeric species. Higher-order oligomerization of SPOP stimulates CRL3SPOP ubiquitination efficiency for its physiological substrate Gli3, suggesting that nuclear speckles are hotspots of ubiquitination. Dynamic, higher-order protein self-association may be a general mechanism to concentrate functional components in membrane-less cellular bodies.",

keywords = "isodesmic self-association, membrane-less organelle, prostate cancer, speckle-type POZ protein, ubiquitin ligase",

author = "Marzahn, {Melissa R.} and Suresh Marada and Jihun Lee and Amanda Nourse and Sophia Kenrick and Huaying Zhao and Gili Ben-Nissan and Kolaitis, {Regina Maria} and Peters, {Jennifer L.} and Stanley Pounds and Errington, {Wesley J.} and Priv{\'e}, {Gilbert G.} and Taylor, {J. Paul} and Michal Sharon and Peter Schuck and Ogden, {Stacey K.} and Tanja Mittag",

note = "Funding Information: We thank BA Schulman, WK Pierce, EW Martin, and CR Grace for valuable discussions and N Clark for preliminary data collection. We also thank H Haecker (St. Jude Children's Research Hospital). This work was funded by a V Foundation Scholar Grant (T.M.), R01GM112846 (T.M.), R01GM101087 (S.K.O.), the National Cancer Institute Cancer Center Support Grant P30CA21765 (at St. Jude Children's Research Hospital), the American Lebanese Syrian Associated Charities (to J.P.T., S.K.O., and T.M.), and the Intramural Program of the National Institute of Biomedical Imaging and Bioengineering (P.S.). Microscopy images were acquired at the Cell & Tissue Imaging Center at St. Jude Children's Research Hospital. Publisher Copyright: {\textcopyright} 2016 The Authors. Published under the terms of the CC BY NC ND 4.0 license",

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month = jun,

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doi = "10.15252/embj.201593169",

language = "English (US)",

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T1 - Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles

AU - Marzahn, Melissa R.

AU - Marada, Suresh

AU - Lee, Jihun

AU - Nourse, Amanda

AU - Kenrick, Sophia

AU - Zhao, Huaying

AU - Ben-Nissan, Gili

AU - Kolaitis, Regina Maria

AU - Peters, Jennifer L.

AU - Pounds, Stanley

AU - Errington, Wesley J.

AU - Privé, Gilbert G.

AU - Taylor, J. Paul

AU - Sharon, Michal

AU - Schuck, Peter

AU - Ogden, Stacey K.

AU - Mittag, Tanja

N1 - Funding Information: We thank BA Schulman, WK Pierce, EW Martin, and CR Grace for valuable discussions and N Clark for preliminary data collection. We also thank H Haecker (St. Jude Children's Research Hospital). This work was funded by a V Foundation Scholar Grant (T.M.), R01GM112846 (T.M.), R01GM101087 (S.K.O.), the National Cancer Institute Cancer Center Support Grant P30CA21765 (at St. Jude Children's Research Hospital), the American Lebanese Syrian Associated Charities (to J.P.T., S.K.O., and T.M.), and the Intramural Program of the National Institute of Biomedical Imaging and Bioengineering (P.S.). Microscopy images were acquired at the Cell & Tissue Imaging Center at St. Jude Children's Research Hospital. Publisher Copyright: © 2016 The Authors. Published under the terms of the CC BY NC ND 4.0 license

PY - 2016/6/15

Y1 - 2016/6/15

N2 - Membrane-less organelles in cells are large, dynamic protein/protein or protein/RNA assemblies that have been reported in some cases to have liquid droplet properties. However, the molecular interactions underlying the recruitment of components are not well understood. Herein, we study how the ability to form higher-order assemblies influences the recruitment of the speckle-type POZ protein (SPOP) to nuclear speckles. SPOP, a cullin-3-RING ubiquitin ligase (CRL3) substrate adaptor, self-associates into higher-order oligomers; that is, the number of monomers in an oligomer is broadly distributed and can be large. While wild-type SPOP localizes to liquid nuclear speckles, self-association-deficient SPOP mutants have a diffuse distribution in the nucleus. SPOP oligomerizes through its BTB and BACK domains. We show that BTB-mediated SPOP dimers form linear oligomers via BACK domain dimerization, and we determine the concentration-dependent populations of the resulting oligomeric species. Higher-order oligomerization of SPOP stimulates CRL3SPOP ubiquitination efficiency for its physiological substrate Gli3, suggesting that nuclear speckles are hotspots of ubiquitination. Dynamic, higher-order protein self-association may be a general mechanism to concentrate functional components in membrane-less cellular bodies.

AB - Membrane-less organelles in cells are large, dynamic protein/protein or protein/RNA assemblies that have been reported in some cases to have liquid droplet properties. However, the molecular interactions underlying the recruitment of components are not well understood. Herein, we study how the ability to form higher-order assemblies influences the recruitment of the speckle-type POZ protein (SPOP) to nuclear speckles. SPOP, a cullin-3-RING ubiquitin ligase (CRL3) substrate adaptor, self-associates into higher-order oligomers; that is, the number of monomers in an oligomer is broadly distributed and can be large. While wild-type SPOP localizes to liquid nuclear speckles, self-association-deficient SPOP mutants have a diffuse distribution in the nucleus. SPOP oligomerizes through its BTB and BACK domains. We show that BTB-mediated SPOP dimers form linear oligomers via BACK domain dimerization, and we determine the concentration-dependent populations of the resulting oligomeric species. Higher-order oligomerization of SPOP stimulates CRL3SPOP ubiquitination efficiency for its physiological substrate Gli3, suggesting that nuclear speckles are hotspots of ubiquitination. Dynamic, higher-order protein self-association may be a general mechanism to concentrate functional components in membrane-less cellular bodies.

KW - isodesmic self-association

KW - membrane-less organelle

KW - prostate cancer

KW - speckle-type POZ protein

KW - ubiquitin ligase

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C2 - 27220849

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SN - 0261-4189

VL - 35

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JO - EMBO Journal

JF - EMBO Journal

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ER -

Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles

Abstract

Bibliographical note

Keywords

UN SDGs

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