| Autor: |
Alvarado-Marchena L; Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Avda. Ingeniero Fausto Elio, 46022 Valencia, Spain., Martínez-Pérez M; Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Avda. Ingeniero Fausto Elio, 46022 Valencia, Spain., Úbeda JR; Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Avda. Ingeniero Fausto Elio, 46022 Valencia, Spain.; Centro de Edafología y Biología Aplicada del Segura (CEBAS)-CSIC, Departamento de Biología del Estrés y Patología Vegetal, 30100 Murcia, Spain., Pallas V; Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Avda. Ingeniero Fausto Elio, 46022 Valencia, Spain., Aparicio F; Instituto de Biología Molecular y Celular de Plantas, Consejo Superior de Investigaciones Científicas-Universitat Politècnica de València, Avda. Ingeniero Fausto Elio, 46022 Valencia, Spain. |
| Abstrakt: |
We have previously reported the presence of m 6 A in the AMV (Alfamovirus, Bromoviridae ) genome. Interestingly, two of these putative m 6 A-sites are in hairpin (hp) structures in the 3'UTR of the viral RNA3. One site ( 2012 AAACU 2016 ) is in the loop of hpB, within the coat protein binding site 1 (CPB1), while the other ( 1900 UGACC 1904 ) is in the lower stem of hpE, a loop previously associated with AMV negative-strand RNA synthesis. In this work, we have performed in vivo experiments to assess the role of these two regions, containing the putative m 6 A-sites in the AMV cycle, by introducing compensatory point mutations to interfere with or abolish the m 6 A-tag of these sites. Our results suggest that the loop of hpB could be involved in viral replication/accumulation. Meanwhile, in the 1900 UGACC 1904 motif of the hpE, the maintenance of the adenosine residue and the lower stem hpE structure are necessary for in vivo plus-strand accumulation. These results extend our understanding of the requirements for hpE in the AMV infection cycle, indicating that both the residue identity and the base-pairing capacity in this structure are essential for viral accumulation. |
