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Extensión no canónica de 3'-5' de ARN con fosfatos ribonucleósidos 2',3'-cíclicos prebióticamente plausibles
Hannes Mutschler1, Philipp Holliger
1MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus , Francis Crick Avenue, Cambridge CB2 0QH, United Kingdom.
Journal of the American Chemical Society
|March 26, 2014
Resumen
Los investigadores diseñaron una ribozima para crear una ruta enzimática de síntesis de ARN utilizando el ribonucleósido 2.
Área de la Ciencia:
- El origen de la vida estudios de estudios.
- La bioquímica del ARN en el ARN.
- Química de los prebióticos.
Sus antecedentes:
- Los fosfatos ribonucleósidos 2',3'-cíclicos (N>p's) son plausibles bloques de construcción prebióticos para el ARN.
- La polimerización no enzimática de N>p en ARN es ineficiente y carece de regioselectividad.
- Anteriormente no se había descrito una ruta enzimática para la polimerización N>p.
Objetivo del estudio:
- Establecer un método enzimático para la síntesis de ARN a partir de N>p's.
- Para investigar el potencial de los ribozimas de ingeniería en el ensamblaje de ARN prebiótico.
Principales métodos:
- Diseñó una variante de la ribozima de la horquilla para la actividad de la nucleotidil transferasa.
- Se utilizó la formación de la fase de hielo eutectico a -7 °C para mejorar la adición de nucleótidos.
- Se probó la adición de los cuatro oligómeros N>p, NAD>p y ARN.
Principales resultados:
- Se ha demostrado actividad de la nucleotidiltransferasa 3'-5' no canónica con los cuatro N>p's.
- Se observó una adición mejorada de nucleótidos 5' facilitada por la formación de hielo eutectico.
- Mostró la adición de dinucleótidos activados y oligómeros de ARN.
Conclusiones:
- Se estableció una nueva vía enzimática para la extensión del ARN 3'-5' utilizando N>p's.
- Destaca el potencial de los ribozimas de ingeniería en la síntesis de ARN de las piscinas prebióticas.
- Sugiere implicaciones para el surgimiento de la vida basada en ARN.
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