Exploración de la interacción entre la región 5´UTR del Sugarcane Mosaic Virus (SCMV) y proteínas del hospedero maíz

Exploración de la interacción entre la región 5´UTR del Sugarcane Mosaic Virus (SCMV) y proteínas del hospedero maíz

Contenido principal del artículo

Giovanni Chaves-Bedoya

Resumen

 

Antecedentes: El virus del mosaico de la caña de azúcar, SCMV (Potyviridae) es un miembro de la gran familia de virus de ARN de cadena positiva, sin una estructura CAP en su extremo 5´no traducido (5`UTR), pero con una proteína viral unida al genoma (VPg) y una cola poli A en su extremo 3´UTR. Se ha sugerido que proteínas del hospedero hacen un puente entre las regiones no traducidas virales 5´y 3´ para potenciar la traducción viral. Dado que las regiones no traducidas presentes en los genomas virales contienen elementos involucrados en la regulación de su ciclo replicativo, es importante analizar la interacción entre estas regiones y las proteínas virales o del hospedero para inferir su función. Objetivo: Determinar si existen proteínas del hospedero maíz que pudieran estar interactuando con la región 5´UTR de SCMV. Metodología: La región 5´UTR de SCMV se amplificó por PCR incluyendo la secuencia del promotor T7 en la secuencia del oligo 5´ UTR y se generaron sus transcritos marcados radiactivamente. Los transcritos marcados fueron entrecruzados con extractos proteicos de hojas de maíz sanas e infectadas. Resultados: Los resultados sugieren la presencia de una proteína putativa de interacción del hospedante maíz de aproximadamente 53kDa, con la región 5´UTR de SCMV. Conclusión: Los ensayos de entrecruzamiento, especificidad y estabilidad de los complejos ARN-proteína sugieren que en el hospedante maíz existe al menos una proteína que interactúa con la región 5´ UTR de SCMV.

Palabras Clave: Complejo ARN-proteína, entrecruzamiento UV, fracción proteica, Potyvirus, VPg.

 

Abstract


Background: Sugarcane mosaic virus is a member of the great family of positive sense RNA viruses, without a CAP structure in its 5´UTR end, but with a viral protein attached to the genome (VPg) and a poli A tail in its 3´UTR end. It has been suggested that some host proteins make a bridge between the untranslated 5´and 3´ regions (UTRs) to enhance the canonical and/or non-canonical virus translation. Since the UTR regions present in the viral genomes have some elements involved in the regulation of their replicative cycle, it is important to analyze the interaction that may occur between these regions and the viral or host proteins to infer regarding its function. Objective: To identify proteins of the host maize that could be interacting with the 5´UTR region of SCMV. Methodology: 5´UTR region of SCMV was amplified by PCR including the sequence of the T7 promoter in the 5´oligo sequence. The transcripts were radioactively labeled. Labeled transcripts were crosslinked with proteins extracts from healthy and infected maize leaves. Results: The results suggest the presence of a 53 kDa putative protein interacting with the 5 ´UTR region of SCMV. Conclusion: Crosslinking essays, specificity and the stability of the RNA-protein complex suggest that in maize host there is at least one protein that interacts with the 5´UTR region of SCMV.

Keywords: RNA-Protein complex, UV crosslinking, protein fraction, Potyvirus, VPg.

 

Resumo


Antecedentes: O vírus do mosaico da cana de açúcar (SCMV) da Família Potyviridae é um membro da grande família de vírus de ARN de cadeia positiva, sem uma estrutura CAP em seu extremo 5´ não traduzido (5`UTR), mas com uma proteína viral unida ao genoma (VPg) e uma cola poli A em seu extremo 3´UTR. Tem-se sugerido que proteínas do hospedeiro fazem uma ponte entre as regiões não traduzidas virales 5´e 3´ para potenciar a tradução viral. Devido que as regiões não traduzidas presentes nos genomas virais contém elementos involucrados na regulação de seu ciclo replicativo, é importante analisar a interação entre estas regiões e as proteínas virales O do hospedeiro para inferir sua função. Objetivo: Determinar se existem proteínas hospedeiras milho que puderam estar interatuando com a região 5´UTR de SCMV. Metodologia: A região 5´UTR de SCMV se amplificou por PCR incluindo a sequência do promotor T7 na sequência do oligo 5´ UTR e se geraram seus transcritos marcados radiativamente. Os transcritos marcados foram entrecruzados com extratos proteicos de folhas de milho saudáveis e infetadas. Resultados: Os resultados sugerem a presença de uma proteína putativa de interação do hospedeiro milho de aproximadamente 53 kDa, com a região 5´UTR de SCMV. Conclusão: Os ensaios de entrecruzamento, especificidade e estabilidade dos complexos ARN proteía sugerem que no hospedeiro milho existe pelo menos uma proteína que interatua com a região 5´ UTR de SCMV.

Palavras-chave: complexo ARN-proteína, entrecruzamento UV, fracção proteica, Potyvirus, VPg.

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Detalles del artículo

Biografía del autor/a (VER)

Giovanni Chaves-Bedoya, Universidad Francisco de Paula Santander

Doctorado en Biotecnología Vegetal. Grupo de Investigación PLANTAE, Facultad de Ciencias Básicas.

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