title: A role for the GSG domain in localizing Sam68 to novel nuclear structures in cancer cell lines.
abstract: The GSG (GRP33, Sam68, GLD-1) domain is a protein module found in an expanding family of RNA-binding proteins. The numerous missense mutations identified genetically in the GSG domain support its physiological role. Although the exact function of the GSG domain is not known, it has been shown to be required for RNA binding and oligomerization. Here it is shown that the Sam68 GSG domain plays a role in protein localization. We show that Sam68 concentrates into novel nuclear structures that are predominantly found in transformed cells. These Sam68 nuclear bodies (SNBs) are distinct from coiled bodies, gems, and promyelocytic nuclear bodies. Electron microscopic studies show that SNBs are distinct structures that are enriched in phosphorus and nitrogen, indicating the presence of nucleic acids. A GFP-Sam68 fusion protein had a similar localization as endogenous Sam68 in HeLa cells, diffusely nuclear with two to five SNBs. Two other GSG proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2, colocalized with endogenous Sam68 in SNBs. Different GSG domain missense mutations were investigated for Sam68 protein localization. Six separate classes of cellular patterns were obtained, including exclusive SNB localization and association with microtubules. These findings demonstrate that the GSG domain is involved in protein localization and define a new compartment for Sam68, SLM-1, and SLM-2 in cancer cell lines.
authors.affiliation: Terry Fox Molecular Oncology Group, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, and Departments of Oncology, Medicine, and Microbiology and Immunology, McGill University, Montréal, Québec H3T 1E2,
authors.forename: T
authors.initials: T
authors.lastname: Chen
authors.forename: F M
authors.initials: FM
authors.lastname: Boisvert
authors.forename: D P
authors.initials: DP
authors.lastname: Bazett-Jones
authors.forename: S
authors.initials: S
authors.lastname: Richard
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D014176:Protein Biosynthesis; D012313:RNA / Q000032:analysis / Q000235:genetics; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000032:analysis / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; C489559:KHDRBS1 protein, human; C485572:KHDRBS2 protein, human; C118777:KHDRBS3 protein, human; C513253:Khdrbs1 protein, mouse; C118776:Khdrbs2 protein, mouse; C486528:Khdrbs3 protein, mouse; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D017411:Ribonucleoproteins, Small Nuclear; D012313:RNA