The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex
The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac12 gene, which is conserved in ten other baculovirus, codes a predicted 217 amino acid protein of unknown function. In this study, we investigated the role of ac12 during baculovirus infection, by generating an ac12 knockout virus....
| Autores principales: | , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2019
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/4491 https://www.sciencedirect.com/science/article/abs/pii/S0168170218304775?via%3Dihub https://doi.org/10.1016/j.virusres.2018.11.010 |
| _version_ | 1855483443035504640 |
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| author | Costa Navarro, Guadalupe Soledad Amalfi, Sabrina Lopez, Maria Gabriela Llauger, Gabriela Arneodo Larochette, Joel Demian Taboga, Oscar Alberto Alfonso, Victoria |
| author_browse | Alfonso, Victoria Amalfi, Sabrina Arneodo Larochette, Joel Demian Costa Navarro, Guadalupe Soledad Llauger, Gabriela Lopez, Maria Gabriela Taboga, Oscar Alberto |
| author_facet | Costa Navarro, Guadalupe Soledad Amalfi, Sabrina Lopez, Maria Gabriela Llauger, Gabriela Arneodo Larochette, Joel Demian Taboga, Oscar Alberto Alfonso, Victoria |
| author_sort | Costa Navarro, Guadalupe Soledad |
| collection | INTA Digital |
| description | The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac12 gene, which is conserved in ten other baculovirus, codes a predicted 217 amino acid protein of unknown function. In this study, we investigated the role of ac12 during baculovirus infection, by generating an ac12 knockout virus. The transfection of the recombinant genome in insect cells resulted in unaltered viral dispersion and occlusion body production when compared to the control bacmid. This finding demonstrates that ac12 is a non-essential gene. Transmission and scanning electron microscopy (SEM) analyses showed that ac12 knockout virus produced occlusion bodies morphologically similar to those obtained with the control and capable to occlude virions. However, a slight but significant size difference was detected by SEM observation of purified occlusion bodies. This difference suggests that ac12 may be involved in regulatory pathways of polyhedrin production or occlusion body assembly without affecting either viral occlusion or oral infectivity in Rachiplusia nu larvae. This was evidenced by bioassays that showed no significant differences in the conditions tested. A qPCR analysis of viral gene expression during infection evidenced regulatory effects of ac12 over some representative genes of different stages of the viral cycle. In this study, we also showed that ac12 is transcribed at early times after infection and remains detectable up to 72 hours post-infection. The mRNA is translated during the infection and results in a protein that encodes an F-box domain that interacts in vivo and in vitro with S phase kinase associated protein 1 (SKP1) adaptor protein, which is potentially involved in protein ubiquitination pathways. |
| format | Artículo |
| id | INTA4491 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA44912019-02-25T18:51:01Z The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex Costa Navarro, Guadalupe Soledad Amalfi, Sabrina Lopez, Maria Gabriela Llauger, Gabriela Arneodo Larochette, Joel Demian Taboga, Oscar Alberto Alfonso, Victoria Autographa Californica Baculovirus Proteínas Virales Viral Proteins AcMNPV Ac12 F-box The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ac12 gene, which is conserved in ten other baculovirus, codes a predicted 217 amino acid protein of unknown function. In this study, we investigated the role of ac12 during baculovirus infection, by generating an ac12 knockout virus. The transfection of the recombinant genome in insect cells resulted in unaltered viral dispersion and occlusion body production when compared to the control bacmid. This finding demonstrates that ac12 is a non-essential gene. Transmission and scanning electron microscopy (SEM) analyses showed that ac12 knockout virus produced occlusion bodies morphologically similar to those obtained with the control and capable to occlude virions. However, a slight but significant size difference was detected by SEM observation of purified occlusion bodies. This difference suggests that ac12 may be involved in regulatory pathways of polyhedrin production or occlusion body assembly without affecting either viral occlusion or oral infectivity in Rachiplusia nu larvae. This was evidenced by bioassays that showed no significant differences in the conditions tested. A qPCR analysis of viral gene expression during infection evidenced regulatory effects of ac12 over some representative genes of different stages of the viral cycle. In this study, we also showed that ac12 is transcribed at early times after infection and remains detectable up to 72 hours post-infection. The mRNA is translated during the infection and results in a protein that encodes an F-box domain that interacts in vivo and in vitro with S phase kinase associated protein 1 (SKP1) adaptor protein, which is potentially involved in protein ubiquitination pathways. Instituto de Biotecnología Fil: Costa Navarro, Guadalupe Soledad. Fundación Instituto Leloir; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Amalfi, Sabrina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Llauger, Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2019-02-25T18:47:13Z 2019-02-25T18:47:13Z 2019-01 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/4491 https://www.sciencedirect.com/science/article/abs/pii/S0168170218304775?via%3Dihub 0168-1702 https://doi.org/10.1016/j.virusres.2018.11.010 eng info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Virus research 260 : 67-77. (15 January 2019) |
| spellingShingle | Autographa Californica Baculovirus Proteínas Virales Viral Proteins AcMNPV Ac12 F-box Costa Navarro, Guadalupe Soledad Amalfi, Sabrina Lopez, Maria Gabriela Llauger, Gabriela Arneodo Larochette, Joel Demian Taboga, Oscar Alberto Alfonso, Victoria The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title | The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title_full | The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title_fullStr | The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title_full_unstemmed | The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title_short | The autographa californica multiple nucleopolyhedrovirus Ac12: A non-essential F box-like protein that interacts with cellular SKP1 component of the E3 ubiquitin ligase complex |
| title_sort | autographa californica multiple nucleopolyhedrovirus ac12 a non essential f box like protein that interacts with cellular skp1 component of the e3 ubiquitin ligase complex |
| topic | Autographa Californica Baculovirus Proteínas Virales Viral Proteins AcMNPV Ac12 F-box |
| url | http://hdl.handle.net/20.500.12123/4491 https://www.sciencedirect.com/science/article/abs/pii/S0168170218304775?via%3Dihub https://doi.org/10.1016/j.virusres.2018.11.010 |
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