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El ARN no codificante del EBV se une al ARN naciente para conducir el PAX5 del huésped al ADN viral
Nara Lee1, Walter N Moss1, Therese A Yario1
1Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University School of Medicine, 295 Congress Avenue, New Haven, CT 06536, USA.
Cell
|February 10, 2015
Resumen
El ARN EBER2 del virus Epstein-Barr guía el factor de transcripción PAX5 hacia el ADN viral. Esta interacción es crucial para regular la expresión y replicación de genes virales, revelando una nueva función para el ARN no codificante.
Área de la Ciencia:
- Virología molecular Virología molecular.
- La epigenética es la epigenética.
- ARN no codificante Biología Biología.
Sus antecedentes:
- El virus de Epstein-Barr (EBV) expresa abundante ARN no codificante nuclear, EBER2.
- La función y localización de EBER2 dentro del núcleo de la célula huésped no estaban claras anteriormente.
- Las repeticiones terminales (TR) del genoma del EBV son regiones reguladoras críticas.
Objetivo del estudio:
- Para investigar la localización de la cromatina del ARN EBER2 EBV EBER2.
- Para dilucidar la interacción funcional entre EBER2 y el factor de transcripción PAX5.5.
- Comprender el papel de la interacción EBER2-PAX5 en la regulación y replicación del gen EBV.
Principales métodos:
- La cromatina, el ARN y la inmunoprecipitación sensible a la temperatura (CHART) para determinar la ubicación genómica de EBER2.
- Inmunoprecipitación del ARN (RIP) para confirmar la interacción EBER2-PAX5.
- Interferencia de ARN (RNAi) para evaluar el impacto funcional de EBER2 knockdown en la expresión y replicación de genes virales.
- Análisis de transcripciones nacientes del locus TR para las interacciones de emparejamiento de bases.
Principales resultados:
- CHART reveló que EBER2 se localiza en el EBV TRs, superponiendo los sitios de unión PAX5.
- EBER2 interactúa directamente con PAX5 y es esencial para su reclutamiento en el TRs.
- El knockdown de EBER2 imita el agotamiento de PAX5, regulando al alza la expresión de LMP2A/B y LMP1 y disminuyendo la replicación lítica.
- El reclutamiento está mediado por el emparejamiento de bases EBER2 con las transcripciones TR nacientes, un mecanismo evolucionariamente conservado.
Conclusiones:
- EBER2 actúa como un andamio, utilizando el emparejamiento de bases con el ARN naciente para reclutar PAX5 a las TRs EBV.
- Este nuevo mecanismo guiado por ARN regula los genes clave EBV latentes y líticos.
- Los hallazgos revelan una función no descrita previamente para el ARN no codificante de acción trans en la orientación de los factores de transcripción a los objetivos del ADN.
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