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A novel assay provides insight into tRNAPhe retrograde nuclear import and re-export in S. cerevisiae.
Nostramo, Regina T; Hopper, Anita K.
Afiliación
  • Nostramo RT; Department of Molecular Genetics Center for RNA Biology The Ohio State University, Columbus, OH 43210, USA.
  • Hopper AK; Department of Molecular Genetics Center for RNA Biology The Ohio State University, Columbus, OH 43210, USA.
Nucleic Acids Res ; 48(20): 11577-11588, 2020 11 18.
Article en En | MEDLINE | ID: mdl-33074312
ABSTRACT
In eukaryotes, tRNAs are transcribed in the nucleus and subsequently exported to the cytoplasm where they serve as essential adaptor molecules in translation. However, tRNAs can be returned to the nucleus by the evolutionarily conserved process called tRNA retrograde nuclear import, before relocalization back to the cytoplasm via a nuclear re-export step. Several important functions of these latter two trafficking events have been identified, yet the pathways are largely unknown. Therefore, we developed an assay in Saccharomyces cerevisiae to identify proteins mediating tRNA retrograde nuclear import and re-export using the unique wybutosine modification of mature tRNAPhe. Our hydrochloric acid/aniline assay revealed that the karyopherin Mtr10 mediates retrograde import of tRNAPhe, constitutively and in response to amino acid deprivation, whereas the Hsp70 protein Ssa2 mediates import specifically in the latter. Furthermore, tRNAPhe is re-exported by Crm1 and Mex67, but not by the canonical tRNA exporters Los1 or Msn5. These findings indicate that the re-export process occurs in a tRNA family-specific manner. Together, this assay provides insights into the pathways for tRNAPhe retrograde import and re-export and is a tool that can be used on a genome-wide level to identify additional gene products involved in these tRNA trafficking events.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: ARN de Transferencia de Fenilalanina / Núcleo Celular / Proteínas de Transporte Nucleocitoplasmático Tipo de estudio: Prognostic_studies Idioma: En Revista: Nucleic Acids Res Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: ARN de Transferencia de Fenilalanina / Núcleo Celular / Proteínas de Transporte Nucleocitoplasmático Tipo de estudio: Prognostic_studies Idioma: En Revista: Nucleic Acids Res Año: 2020 Tipo del documento: Article País de afiliación: Estados Unidos