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[5´cap -independent translation of dengue virus genomic RNA]. / Traducción independiente de la estructura 5´cap del ARN genómico del virus dengue.
Pérez Dominguez, Mariela; Rojas, Roselbis; Bandeira, Eduardo; Requena, Dayana; Ferreras, Ana C; Triana, Juana L; Triana-Alonso, Francisco.
Afiliação
  • Pérez Dominguez M; Facultad de Odontología, Universidad de Carabobo, Valencia, Venezuela.
  • Rojas R; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
  • Bandeira E; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
  • Requena D; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
  • Ferreras AC; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
  • Triana JL; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
  • Triana-Alonso F; Facultad de Ciencias de la Salud, Universidad de Carabobo, Maracay, Venezuela.
Rev Peru Med Exp Salud Publica ; 32(1): 11-8, 2015.
Article em Es | MEDLINE | ID: mdl-26102100
OBJETIVES: To analyze the involvement of methyl guanosine triphosphate cap (5’cap) and the start site of the genomic RNA of Dengue virus serotype 2 (DENV-2) American genotype in translation, using a cell-free system prepared from human placenta. MATERIALS AND METHODS: The recombinant plasmid pTZ18R-D2 was prepared containing DNA encoding the 5’UTR and the first 201 nucleotides of the viral capsid. This plasmid was used to transcribe the corresponding RNA (RNA-D2) without the 5’ cap. The RNA-D2 was translated in a system consisting of the postmitochondrial fraction (S-30) from human placenta and the incorporation of [14C] aminoacids in the presence of RNA-D2 and in its absence (control) was evaluated. Seven antisense oligonucleotides (OAs1-7) directed against sequences of the SLA, SLB and CHP structures of RNA-D2 were designed and the effect thereof on RNA-D2 translation was analyzed. RESULTS: The RNA-D2 produced a significant increase (p<0.001) in the incorporation of [14C] amino acids, with 75% stimulation of translational activity compared to the control. Analysis of the translation products showed peak incorporation corresponding to peptides with apparent molecular weight close to the expected (7.746 kDa).The OAs5, complementary to a sequence of SLB structure of RNA-D2, completely inhibited translation. CONCLUSIONS: The RNA-D2 was translated specifically and efficiently under conditions similar to human intracellular conditions, by an alternative 5’ cap-independent mechanism, which would involve the SLB structure. This mechanism might be seen as an aim in the development of antisense therapies to inhibit virus replication.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Biossíntese de Proteínas / Capuzes de RNA / RNA Viral / Vírus da Dengue Limite: Humans Idioma: Es Revista: Rev Peru Med Exp Salud Publica Assunto da revista: SAUDE PUBLICA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: Venezuela País de publicação: Peru
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Biossíntese de Proteínas / Capuzes de RNA / RNA Viral / Vírus da Dengue Limite: Humans Idioma: Es Revista: Rev Peru Med Exp Salud Publica Assunto da revista: SAUDE PUBLICA Ano de publicação: 2015 Tipo de documento: Article País de afiliação: Venezuela País de publicação: Peru