Comparative ultrastructure of CRM1-Nucleolar bodies (CNoBs), Intranucleolar bodies (INBs) and hybrid PML/p62 bodies uncovers new facets of nuclear body dynamic and diversity.
In order to gain insights on the nuclear organization in mammalian cells, we characterized ultrastructurally nuclear bodies (NBs) previously described as fluorescent foci. Using high resolution immunoelectron microscopy (I-EM), we provide evidence that CNoBs (CRM1-Nucleolar bodies) and INBs (Intranucleolar bodies) are distinct genuine nucleolar structures in untreated HeLa cells. INBs are fibrillar and concentrate the post-translational modifiers SUMO1 and SUMO-2/3 as strongly as PML bodies. In contrast, the smallest CRM1-labeled CNoBs are vitreous, preferentially located at the periphery of the nucleolus and, intricately linked to the chromatin network. Upon blockage of the CRM1-dependent nuclear export by leptomycin B (LMB), CNoBs disappear while p62/SQSTM1-containing fibrillar nuclear bodies are induced. These p62 bodies are enriched in ubiquitinated proteins. They progressively associate with PML bodies to form hybrid bodies of which PML decorates the periphery while p62/SQSTM1 is centrally-located. Our study is expanding the repertoire of nuclear bodies; revealing a previously unrecognized composite nucleolar landscape and a new mode of interactions between ubiquitous (PML) and stress-induced (p62) nuclear bodies, resulting in the formation of hybrid bodies.
a Functional Organization of the Cell ; CNRS UMR-9196 ; Institut Gustave Roussy ; Villejuif , France.
Sylvie
S
Souquere
Dominique
D
Weil
Gérard
G
Pierron
D021581:Active Transport, Cell Nucleus; D002466:Cell Nucleolus / Q000648:ultrastructure; D002843:Chromatin / Q000737:chemistry / Q000235:genetics; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D045586:Intranuclear Inclusion Bodies / Q000235:genetics / Q000648:ultrastructure*; D028884:Karyopherins / Q000235:genetics / Q000648:ultrastructure*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000648:ultrastructure*; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism; D000097606:Exportin 1 Protein
D002843:Chromatin; D005231:Fatty Acids, Unsaturated; D028884:Karyopherins; D018160:Receptors, Cytoplasmic and Nuclear; D025842:SUMO-1 Protein; D000097606:Exportin 1 Protein; C495501:SUMO1 protein, human; C038753:leptomycin B
CNoB; INB; PML bodies; SUMO; immunoelectron microscopy; nuclear bodies; nuclear organization; nucleolus; p62/SQSTM1; ubiquitin