Paraspeckle subnuclear bodies depend on dynamic heterodimerisation of DBHS RNA-binding proteins via their structured domains

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dc.contributor.authorLee, P.W.
dc.contributor.authorMarshall, A.C.
dc.contributor.authorKnott, G.J.
dc.contributor.authorKobelke, S.
dc.contributor.authorMartelotto, L.
dc.contributor.authorCho, E.
dc.contributor.authorMcMillan, P.J.
dc.contributor.authorLee, M.
dc.contributor.authorBond, C.S.
dc.contributor.authorFox, A.H.
dc.date.issued2022
dc.description.abstractRNA-binding proteins of the DBHS (Drosophila Behavior Human Splicing) family, NONO, SFPQ, and PSPC1 have numerous roles in genome stability and transcriptional and posttranscriptional regulation. Critical to DBHS activity is their recruitment to distinct subnuclear locations, for example, paraspeckle condensates, where DBHS proteins bind to the long noncoding RNA NEAT1 in the first essential step in paraspeckle formation. To carry out their diverse roles, DBHS proteins form homodimers and heterodimers, but how this dimerization influences DBHS localization and function is unknown. Here, we present an inducible GFP-NONO stable cell line and use it for live-cell 3D-structured illumination microscopy, revealing paraspeckles with dynamic, twisted elongated structures. Using siRNA knockdowns, we show these labeled paraspeckles consist of GFP-NONO/endogenous SFPQ dimers and that GFP-NONO localization to paraspeckles depends on endogenous SFPQ. Using purified proteins, we confirm that partner swapping between NONO and SFPQ occurs readily in vitro. Crystallographic analysis of the NONOSFPQ heterodimer reveals conformational differences to the other DBHS dimer structures, which may contribute to partner preference, RNA specificity, and subnuclear localization. Thus overall, our study suggests heterodimer partner availability is crucial for NONO subnuclear distribution and helps explain the complexity of both DBHS protein and paraspeckle dynamics through imaging and structural approaches.
dc.description.statementofresponsibilityPei Wen Lee, Andrew C. Marshall, Gavin J. Knott, Simon Kobelke, Luciano Martelotto, Ellie Cho, Paul J. McMillan, Mihwa Lee, Charles S. Bond, and Archa H. Fox
dc.identifier.citationJournal of Biological Chemistry, 2022; 298(11):1-15
dc.identifier.doi10.1016/j.jbc.2022.102563
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.orcidMartelotto, L. [0000-0002-9625-1183]
dc.identifier.urihttps://hdl.handle.net/2440/136996
dc.language.isoen
dc.publisherElsevier BV
dc.relation.granthttp://purl.org/au-research/grants/arc/FT180100204
dc.relation.granthttp://purl.org/au-research/grants/arc/DP160102435
dc.rights© 2022 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
dc.source.urihttps://doi.org/10.1016/j.jbc.2022.102563
dc.subjectdimerization
dc.subjectDrosophila behavior human splicing (DBHS)
dc.subjectimaging
dc.subjectlong noncoding RNA
dc.subjectNONO
dc.subjectnuclear organization
dc.subjectparaspeckles
dc.subjectRNA-binding protein
dc.subjectSFPQ
dc.subjectsuper-resolution microscopy
dc.subject.meshHumans
dc.subject.meshRNA-Binding Proteins
dc.subject.meshGene Expression Regulation
dc.subject.meshDimerization
dc.subject.meshRNA, Long Noncoding
dc.subject.meshParaspeckles
dc.titleParaspeckle subnuclear bodies depend on dynamic heterodimerisation of DBHS RNA-binding proteins via their structured domains
dc.typeJournal article
pubs.publication-statusPublished

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