title: Ty1 gag enhances the stability and nuclear export of Ty1 mRNA.
abstract: Retrotransposon and retroviral RNA delivery to particle assembly sites is essential for their replication. mRNA and Gag from the Ty1 retrotransposon colocalize in cytoplasmic foci, which are required for transposition and may be the sites for virus-like particle (VLP) assembly. To determine which Ty1 components are required to form mRNA/Gag foci, localization studies were performed in a Ty1-less strain expressing galactose-inducible Ty1 plasmids (pGTy1) containing mutations in GAG or POL. Ty1 mRNA/Gag foci remained unaltered in mutants defective in Ty1 protease (PR) or deleted for POL. However, Ty1 mRNA containing a frameshift mutation (Ty1fs) that prevents the synthesis of all proteins accumulated in the nucleus. Ty1fs RNA showed a decrease in stability that was mediated by the cytoplasmic exosome, nonsense-mediated decay (NMD) and the processing body. Localization of Ty1fs RNA remained unchanged in an nmd2Δ mutant. When Gag and Ty1fs mRNA were expressed independently, Gag provided in trans increased Ty1fs RNA level and restored localization of Ty1fs RNA in cytoplasmic foci. Endogenously expressed Gag also localized to the nuclear periphery independent of RNA export. These results suggest that Gag is required for Ty1 mRNA stability, efficient nuclear export and localization into cytoplasmic foci.
authors.affiliation: Gene Regulation and Chromosome Biology Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD 21702, USA.
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Checkley
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Mitchell
authors.forename: Linda D
authors.initials: LD
authors.lastname: Eizenstat
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Lockett
authors.forename: David J
authors.initials: DJ
authors.lastname: Garfinkel
mesh_terms: D021581:Active Transport, Cell Nucleus; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D055354:Exosomes / Q000378:metabolism; D016368:Frameshift Mutation; D015685:Fusion Proteins, gag-pol / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D020871:RNA Stability; D034443:RNA Transport; D012333:RNA, Messenger / Q000378:metabolism*; D018626:Retroelements / Q000235:genetics*; D012190:Retroviridae / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D015685:Fusion Proteins, gag-pol; C092470:NMD2 protein, S cerevisiae; D012333:RNA, Messenger; D018626:Retroelements; D029701:Saccharomyces cerevisiae Proteins