The Membrane-Anchoring Region of the AcMNPV P74 Protein Is Expendable or Interchangeable with Homologs from Other Species
María Victoria Nugnes,
Alexandra Marisa Targovnik,
Adrià Mengual-Martí,
María Victoria Miranda,
Carolina Susana Cerrudo,
Salvador Herrero,
Mariano Nicolás Belaich
Affiliations
María Victoria Nugnes
Laboratorio de Ingeniería Genética y Biología Celular y Molecular—Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, Buenos Aires B1876BXD, Argentina
Alexandra Marisa Targovnik
Cátedra de Biotecnología, Departamento de Microbiología, Inmunología, Biotecnología y Genética, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, Sexto Piso, Buenos Aires C1113AAD, Argentina
Adrià Mengual-Martí
Department of Genetics and University Institute of Biotechnology and Biomedicine-BIOTECMED, Universitat de València, Dr. Moliner 50, Valencia, 46100 Burjassot, Spain
María Victoria Miranda
Cátedra de Biotecnología, Departamento de Microbiología, Inmunología, Biotecnología y Genética, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, Sexto Piso, Buenos Aires C1113AAD, Argentina
Carolina Susana Cerrudo
Laboratorio de Ingeniería Genética y Biología Celular y Molecular—Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, Buenos Aires B1876BXD, Argentina
Salvador Herrero
Department of Genetics and University Institute of Biotechnology and Biomedicine-BIOTECMED, Universitat de València, Dr. Moliner 50, Valencia, 46100 Burjassot, Spain
Mariano Nicolás Belaich
Laboratorio de Ingeniería Genética y Biología Celular y Molecular—Área Virosis de Insectos, Instituto de Microbiología Básica y Aplicada, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Sáenz Peña 352, Bernal, Buenos Aires B1876BXD, Argentina
Baculoviruses are insect pathogens that are characterized by assembling the viral dsDNA into two different enveloped virions during an infective cycle: occluded virions (ODVs; immersed in a protein matrix known as occlusion body) and budded virions (BVs). ODVs are responsible for the primary infection in midgut cells of susceptible larvae thanks to the per os infectivity factor (PIF) complex, composed of at least nine essential viral proteins. Among them, P74 is a crucial factor whose activity has been identified as virus-specific. In this work, the p74 gene from AcMNPV was pseudogenized using CRISPR/Cas9 technology and then complemented with wild-type alleles from SeMNPV and HearSNPV species, as well as chimeras combining the P74 amino and carboxyl domains. The results on Spodoptera exigua and Rachiplusia nu larvae showed that an amino terminal sector of P74 (lacking two potential transmembrane regions but possessing a putative nuclear export signal) is sufficient to restore the virus infectivity whether alone or fused to the P74 transmembrane regions of the other evaluated viral species. These results provide novel information about the functional role of P74 and delimit the region on which mutagenesis could be applied to enhance viral activity and, thus, produce better biopesticides.
