title: Structural, super-resolution microscopy analysis of paraspeckle nuclear body organization.
abstract: Paraspeckles are nuclear bodies built on the long noncoding RNA Neat1, which regulates a variety of physiological processes including cancer progression and corpus luteum formation. To obtain further insight into the molecular basis of the function of paraspeckles, we performed fine structural analyses of these nuclear bodies using structural illumination microscopy. Notably, paraspeckle proteins are found within different layers along the radially arranged bundles of Neat1 transcripts, forming a characteristic core-shell spheroidal structure. In cells lacking the RNA binding protein Fus, paraspeckle spheroids are disassembled into smaller particles containing Neat1, which are diffusely distributed in the nucleoplasm. Sequencing analysis of RNAs purified from paraspeckles revealed that AG-rich transcripts associate with Neat1, which are distributed along the shell of the paraspeckle spheroids. We propose that paraspeckles sequester core components inside the spheroids, whereas the outer surface associates with other components in the nucleoplasm to fulfill their function.
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114 Department of Genetics, Harvard Medical School, Boston, MA 02115.
authors.forename: Jason A
authors.identifier: 0000-0002-7252-8651
authors.initials: JA
authors.lastname: West
authors.affiliation: RNA Biology Laboratory, RIKEN, Wako 351-0198, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Mito
authors.affiliation: RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Satoshi
authors.identifier: 0000-0002-4365-9003
authors.initials: S
authors.lastname: Kurosaka
authors.affiliation: RIKEN Brain Science Institute, Wako, Saitama 351-0198, Japan.
authors.forename: Toru
authors.identifier: 0000-0001-7153-266X
authors.initials: T
authors.lastname: Takumi
authors.affiliation: Phyloinformatics Unit, RIKEN Center for Life Science Technologies, Chuo-ku, Kobe, Hyogo 650-0047, Japan.
authors.forename: Chiharu
authors.identifier: 0000-0003-4621-9027
authors.initials: C
authors.lastname: Tanegashima
authors.affiliation: Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
authors.forename: Takeshi
authors.identifier: 0000-0003-2393-5776
authors.initials: T
authors.lastname: Chujo
authors.affiliation: RNA Biology Laboratory, RIKEN, Wako 351-0198, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Yanaka
authors.affiliation: Department of Molecular Biology, Massachusetts General Hospital, Boston, MA 02114 Department of Genetics, Harvard Medical School, Boston, MA 02115.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Kingston
authors.affiliation: Institute for Genetic Medicine, Hokkaido University, Sapporo 060-0815, Japan.
authors.forename: Tetsuro
authors.identifier: 0000-0003-1068-5464
authors.initials: T
authors.lastname: Hirose
authors.affiliation: School of Chemistry and Biochemistry, The University of Western Australia, Crawley, Western Australia 6009, Australia.
authors.forename: Charles
authors.identifier: 0000-0002-9584-6783
authors.initials: C
authors.lastname: Bond
authors.affiliation: School of Anatomy, Physiology and Human Biology, The University of Western Australia, Crawley, Western Australia 6009, Australia Harry Perkins Institute of Medical Research, Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia.
authors.forename: Archa
authors.identifier: 0000-0003-1962-270X
authors.initials: A
authors.lastname: Fox
authors.affiliation: RNA Biology Laboratory, RIKEN, Wako 351-0198, Japan RNA Biology Laboratory, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812, Japan nakagawas@pharm.hokudai.ac.jp.
authors.forename: Shinichi
authors.identifier: 0000-0003-1697-1234
authors.initials: S
authors.lastname: Nakagawa
mesh_terms: D000818:Animals; D001483:Base Sequence; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D045586:Intranuclear Inclusion Bodies / Q000378:metabolism*; D051379:Mice; D008853:Microscopy / Q000379:methods*; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism; D034702:RNA-Binding Protein FUS / Q000378:metabolism; D017423:Sequence Analysis, RNA
chemical_list: C577275:NEAT1 long non-coding RNA, mouse; D062085:RNA, Long Noncoding; D034702:RNA-Binding Protein FUS