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

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 abili...

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Published in:The EMBO journal 2016-06, Vol.35 (12), p.1254-1275
Main Authors: Marzahn, Melissa R, Marada, Suresh, Lee, Jihun, Nourse, Amanda, Kenrick, Sophia, Zhao, Huaying, Ben-Nissan, Gili, Kolaitis, Regina-Maria, Peters, Jennifer L, Pounds, Stanley, Errington, Wesley J, Privé, Gilbert G, Taylor, J Paul, Sharon, Michal, Schuck, Peter, Ogden, Stacey K, Mittag, Tanja
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cited_by cdi_FETCH-LOGICAL-c5519-b0ac1a895c12acebe4e2c68266e6e8f587f7babbb6f3f8ffb2cd7ff9434edcbd3
cites cdi_FETCH-LOGICAL-c5519-b0ac1a895c12acebe4e2c68266e6e8f587f7babbb6f3f8ffb2cd7ff9434edcbd3
container_end_page 1275
container_issue 12
container_start_page 1254
container_title The EMBO journal
container_volume 35
creator Marzahn, Melissa R
Marada, Suresh
Lee, Jihun
Nourse, Amanda
Kenrick, Sophia
Zhao, Huaying
Ben-Nissan, Gili
Kolaitis, Regina-Maria
Peters, Jennifer L
Pounds, Stanley
Errington, Wesley J
Privé, Gilbert G
Taylor, J Paul
Sharon, Michal
Schuck, Peter
Ogden, Stacey K
Mittag, Tanja
description 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. Synopsis SPOP, a ubiquitin ligase substrate receptor and tumor suppressor, self‐associates indefinitely into large oligomers via the synergistic function of its tandem dimerization domains. The resulting oligomers are recruited to liquid nuclear speckles, likely generating hotspots of SPOP‐mediated ubiquitination. SPOP localizes to liquid nuclear bodies. Self‐association‐deficient SPOP mutants lose their localization to nuclear speckles. SPOP forms labile higher‐order oligomers through tandem self‐association domains and an isodesmic mechanism. The BTB self‐association‐deficient mutant has a dominant‐negative effect on Hedgehog signaling in the developing fly wing. Dynamic, higher‐order self‐association may be a general mechanism to concentrate functional components in membrane‐less cellular bodies. Graphical Abstract Self‐association of the ubiquitin ligase adaptor and tumor suppressor SPOP is required for its recruitment to liquid nuclear bodies, likely generating hotspots of SPOP‐mediated ubiquitination.
doi_str_mv 10.15252/embj.201593169
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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. Synopsis SPOP, a ubiquitin ligase substrate receptor and tumor suppressor, self‐associates indefinitely into large oligomers via the synergistic function of its tandem dimerization domains. The resulting oligomers are recruited to liquid nuclear speckles, likely generating hotspots of SPOP‐mediated ubiquitination. SPOP localizes to liquid nuclear bodies. Self‐association‐deficient SPOP mutants lose their localization to nuclear speckles. SPOP forms labile higher‐order oligomers through tandem self‐association domains and an isodesmic mechanism. The BTB self‐association‐deficient mutant has a dominant‐negative effect on Hedgehog signaling in the developing fly wing. Dynamic, higher‐order self‐association may be a general mechanism to concentrate functional components in membrane‐less cellular bodies. 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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. Synopsis SPOP, a ubiquitin ligase substrate receptor and tumor suppressor, self‐associates indefinitely into large oligomers via the synergistic function of its tandem dimerization domains. The resulting oligomers are recruited to liquid nuclear speckles, likely generating hotspots of SPOP‐mediated ubiquitination. SPOP localizes to liquid nuclear bodies. Self‐association‐deficient SPOP mutants lose their localization to nuclear speckles. SPOP forms labile higher‐order oligomers through tandem self‐association domains and an isodesmic mechanism. 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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. Synopsis SPOP, a ubiquitin ligase substrate receptor and tumor suppressor, self‐associates indefinitely into large oligomers via the synergistic function of its tandem dimerization domains. The resulting oligomers are recruited to liquid nuclear speckles, likely generating hotspots of SPOP‐mediated ubiquitination. SPOP localizes to liquid nuclear bodies. Self‐association‐deficient SPOP mutants lose their localization to nuclear speckles. SPOP forms labile higher‐order oligomers through tandem self‐association domains and an isodesmic mechanism. The BTB self‐association‐deficient mutant has a dominant‐negative effect on Hedgehog signaling in the developing fly wing. Dynamic, higher‐order self‐association may be a general mechanism to concentrate functional components in membrane‐less cellular bodies. Graphical Abstract Self‐association of the ubiquitin ligase adaptor and tumor suppressor SPOP is required for its recruitment to liquid nuclear bodies, likely generating hotspots of SPOP‐mediated ubiquitination.</abstract><cop>London</cop><pub>Blackwell Publishing Ltd</pub><pmid>27220849</pmid><doi>10.15252/embj.201593169</doi><tpages>22</tpages><orcidid>https://orcid.org/0000-0002-1827-3811</orcidid><oa>free_for_read</oa></addata></record>
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subjects Cell Nucleus - metabolism
EMBO31
EMBO32
EMBO40
Humans
isodesmic self-association
Kruppel-Like Transcription Factors - metabolism
Localization
Macromolecular Substances - metabolism
membrane-less organelle
Nerve Tissue Proteins - metabolism
Nuclear Proteins - metabolism
prostate cancer
Protein Binding
Protein Domains
Protein Multimerization
Proteins
Repressor Proteins - metabolism
speckle-type POZ protein
ubiquitin ligase
Ubiquitination
Zinc Finger Protein Gli3
title Higher-order oligomerization promotes localization of SPOP to liquid nuclear speckles
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