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A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae

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 relocaliz...

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Autores principales: Nostramo, Regina T, Hopper, Anita K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672469/
https://www.ncbi.nlm.nih.gov/pubmed/33074312
http://dx.doi.org/10.1093/nar/gkaa879
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author Nostramo, Regina T
Hopper, Anita K
author_facet Nostramo, Regina T
Hopper, Anita K
author_sort Nostramo, Regina T
collection PubMed
description 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 tRNA(Phe). Our hydrochloric acid/aniline assay revealed that the karyopherin Mtr10 mediates retrograde import of tRNA(Phe), constitutively and in response to amino acid deprivation, whereas the Hsp70 protein Ssa2 mediates import specifically in the latter. Furthermore, tRNA(Phe) 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 tRNA(Phe) 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.
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spelling pubmed-76724692020-11-24 A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae Nostramo, Regina T Hopper, Anita K Nucleic Acids Res Molecular Biology 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 tRNA(Phe). Our hydrochloric acid/aniline assay revealed that the karyopherin Mtr10 mediates retrograde import of tRNA(Phe), constitutively and in response to amino acid deprivation, whereas the Hsp70 protein Ssa2 mediates import specifically in the latter. Furthermore, tRNA(Phe) 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 tRNA(Phe) 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. Oxford University Press 2020-10-19 /pmc/articles/PMC7672469/ /pubmed/33074312 http://dx.doi.org/10.1093/nar/gkaa879 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Molecular Biology
Nostramo, Regina T
Hopper, Anita K
A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title_full A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title_fullStr A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title_full_unstemmed A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title_short A novel assay provides insight into tRNA(Phe) retrograde nuclear import and re-export in S. cerevisiae
title_sort novel assay provides insight into trna(phe) retrograde nuclear import and re-export in s. cerevisiae
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672469/
https://www.ncbi.nlm.nih.gov/pubmed/33074312
http://dx.doi.org/10.1093/nar/gkaa879
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