Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity
The RNA3 species of the beet necrotic yellow vein virus (BNYVV), a multipartite positive-stranded RNA phytovirus, contains the ‘core’ nucleotide sequence required for its systemic movement in Beta macrocarpa. Within this ‘core’ sequence resides a conserved “coremin” motif of 20 nucleotides that is a...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2018-03-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1999-4915/10/3/137 |
| _version_ | 1818417148221980672 |
|---|---|
| author | Alyssa Flobinus Nicolas Chevigny Phillida A. Charley Tanja Seissler Elodie Klein Claudine Bleykasten-Grosshans Claudio Ratti Salah Bouzoubaa Jeffrey Wilusz David Gilmer |
| author_facet | Alyssa Flobinus Nicolas Chevigny Phillida A. Charley Tanja Seissler Elodie Klein Claudine Bleykasten-Grosshans Claudio Ratti Salah Bouzoubaa Jeffrey Wilusz David Gilmer |
| author_sort | Alyssa Flobinus |
| collection | DOAJ |
| description | The RNA3 species of the beet necrotic yellow vein virus (BNYVV), a multipartite positive-stranded RNA phytovirus, contains the ‘core’ nucleotide sequence required for its systemic movement in Beta macrocarpa. Within this ‘core’ sequence resides a conserved “coremin” motif of 20 nucleotides that is absolutely essential for long-distance movement. RNA3 undergoes processing steps to yield a noncoding RNA3 (ncRNA3) possessing “coremin” at its 5′ end, a mandatory element for ncRNA3 accumulation. Expression of wild-type (wt) or mutated RNA3 in Saccharomyces cerevisiae allows for the accumulation of ncRNA3 species. Screening of S. cerevisiae ribonuclease mutants identified the 5′-to-3′ exoribonuclease Xrn1 as a key enzyme in RNA3 processing that was recapitulated both in vitro and in insect cell extracts. Xrn1 stalled on ncRNA3-containing RNA substrates in these decay assays in a similar fashion as the flavivirus Xrn1-resistant structure (sfRNA). Substitution of the BNYVV-RNA3 ‘core’ sequence by the sfRNA sequence led to the accumulation of an ncRNA species in yeast in vitro but not in planta and no viral long distance occurred. Interestingly, XRN4 knockdown reduced BNYVV RNA accumulation suggesting a dual role for the ribonuclease in the viral cycle. |
| first_indexed | 2024-12-14T12:02:10Z |
| format | Article |
| id | doaj.art-765160ae9d774f2b9d57d6dab019a6e7 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-14T12:02:10Z |
| publishDate | 2018-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-765160ae9d774f2b9d57d6dab019a6e72022-12-21T23:01:58ZengMDPI AGViruses1999-49152018-03-0110313710.3390/v10030137v10030137Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease ActivityAlyssa Flobinus0Nicolas Chevigny1Phillida A. Charley2Tanja Seissler3Elodie Klein4Claudine Bleykasten-Grosshans5Claudio Ratti6Salah Bouzoubaa7Jeffrey Wilusz8David Gilmer9Institut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceInstitut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceDepartment of Microbiology, Immunology & Pathology, Colorado State University, Fort Collins, CO 80523-168, USAInstitut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceInstitut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceGénétique moléculaire génomique microbiologie, CNRS UMR7156, 67000 Strasbourg, FranceDipSA-Plant Pathology, University of Bologna, 40127 Bologna, ItalyInstitut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceDepartment of Microbiology, Immunology & Pathology, Colorado State University, Fort Collins, CO 80523-168, USAInstitut de biologie moléculaire des plantes, CNRS UPR2357, Université de Strasbourg, 67084 Strasbourg, FranceThe RNA3 species of the beet necrotic yellow vein virus (BNYVV), a multipartite positive-stranded RNA phytovirus, contains the ‘core’ nucleotide sequence required for its systemic movement in Beta macrocarpa. Within this ‘core’ sequence resides a conserved “coremin” motif of 20 nucleotides that is absolutely essential for long-distance movement. RNA3 undergoes processing steps to yield a noncoding RNA3 (ncRNA3) possessing “coremin” at its 5′ end, a mandatory element for ncRNA3 accumulation. Expression of wild-type (wt) or mutated RNA3 in Saccharomyces cerevisiae allows for the accumulation of ncRNA3 species. Screening of S. cerevisiae ribonuclease mutants identified the 5′-to-3′ exoribonuclease Xrn1 as a key enzyme in RNA3 processing that was recapitulated both in vitro and in insect cell extracts. Xrn1 stalled on ncRNA3-containing RNA substrates in these decay assays in a similar fashion as the flavivirus Xrn1-resistant structure (sfRNA). Substitution of the BNYVV-RNA3 ‘core’ sequence by the sfRNA sequence led to the accumulation of an ncRNA species in yeast in vitro but not in planta and no viral long distance occurred. Interestingly, XRN4 knockdown reduced BNYVV RNA accumulation suggesting a dual role for the ribonuclease in the viral cycle.http://www.mdpi.com/1999-4915/10/3/137Viral noncoding RNAexoribonucleaseflavivirusBNYVVVIGS |
| spellingShingle | Alyssa Flobinus Nicolas Chevigny Phillida A. Charley Tanja Seissler Elodie Klein Claudine Bleykasten-Grosshans Claudio Ratti Salah Bouzoubaa Jeffrey Wilusz David Gilmer Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity Viruses Viral noncoding RNA exoribonuclease flavivirus BNYVV VIGS |
| title | Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity |
| title_full | Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity |
| title_fullStr | Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity |
| title_full_unstemmed | Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity |
| title_short | Beet Necrotic Yellow Vein Virus Noncoding RNA Production Depends on a 5′→3′ Xrn Exoribonuclease Activity |
| title_sort | beet necrotic yellow vein virus noncoding rna production depends on a 5 3 xrn exoribonuclease activity |
| topic | Viral noncoding RNA exoribonuclease flavivirus BNYVV VIGS |
| url | http://www.mdpi.com/1999-4915/10/3/137 |
| work_keys_str_mv | AT alyssaflobinus beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT nicolaschevigny beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT phillidaacharley beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT tanjaseissler beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT elodieklein beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT claudinebleykastengrosshans beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT claudioratti beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT salahbouzoubaa beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT jeffreywilusz beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity AT davidgilmer beetnecroticyellowveinvirusnoncodingrnaproductiondependsona53xrnexoribonucleaseactivity |