title: ASH1 mRNP-core factors form stable complexes in absence of cargo RNA at physiological conditions.
abstract: Asymmetric ASH1 mRNA transport during mitosis of budding yeast constitutes one of the best-studied examples of mRNA localization. Recently, 2 studies used in vitro motility assays to prove that motile ASH1 mRNA-transport complexes can be reconstituted entirely from recombinant factors. Both studies, however, differed in their conclusions on whether cargo RNA itself is required for particle assembly and thus activation of directional transport. Here we provide direct evidence that stable complexes do assemble in absence of RNA at physiologic conditions and even at ionic strengths above cellular levels. These results directly confirm the previous notion that the ASH1 transport machinery is not activated by the cargo RNA itself, but rather through protein-protein interactions.
authors.affiliation: a Institute of Structural Biology ; Helmholtz Zentrum München - German Center for Environmental Health ; Neuherberg , Germany.
authors.forename: Franziska T
authors.initials: FT
authors.lastname: Edelmann
authors.forename: Annika
authors.initials: A
authors.lastname: Niedner
authors.forename: Dierk
authors.initials: D
authors.lastname: Niessing
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008938:Mitosis; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D024701:Myosin Type V / Q000235:genetics / Q000378:metabolism; D009994:Osmolar Concentration; D010766:Phosphorylation; D055503:Protein Multimerization; D034443:RNA Transport; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: C098733:ASH1 protein, S cerevisiae; C098732:Myo4 protein, S cerevisiae; D012331:RNA, Fungal; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D012261:Ribonucleoproteins; C471175:SHE2 protein, S cerevisiae; C471176:SHE3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C026452:messenger ribonucleoprotein; D024701:Myosin Type V; D018995:Myosin Heavy Chains
keywords: ASH1 mRNA; Myo4p; RNA localization; She2p; She3p; budding yeast; in vitro reconstitution; mRNP; macromolecular assembly; myosin
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