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Estructura y termodinámica de la N6-metiladenosina en el ARN: una modificación de la base con muelles

Caroline Roost¹, Stephen R Lynch, Pedro J Batista

¹Department of Chemistry, ‡Howard Hughes Medical Institute and Program in Epithelial Biology, Stanford University , Stanford, California 94305, United States.

Journal of the American Chemical Society

|January 23, 2015

Resumen

La N(6)-metiladenosina (m(6) A) altera la estructura del ARN, desestabilizando las regiones de doble cadena pero estabilizando las de una sola cadena. Esta modificación influye en la estructura secundaria del ARN y en los procesos celulares.

Área de la Ciencia:

Biología Molecular Biología Molecular
La bioquímica es la bioquímica.
ARN Biología Biología ARN

Sus antecedentes:

La N(6)-metiladenosina (m(6) A) es una modificación prevalente del ARN.
m(6) A está relacionado con la degradación, localización y empalme del ARN.
Los mecanismos moleculares precisos de la función de m(6) A siguen sin estar claros.

Objetivo del estudio:

Para aclarar el impacto estructural y termodinámico de las modificaciones m(6) A en el ARN.
Para investigar cómo m(6) A influye en la estructura secundaria del ARN en un contexto celular.

Principales métodos:

Espectroscopia de RMN 2D para determinar las estructuras dúplex de ARN con y sin m6A.
Mediciones termodinámicas de m(6) A en duplexos de ARN.
Mapeo de nucleasas en todo el transcriptoma para analizar la estructura secundaria de ARN metilado en células humanas.

Principales resultados:

m(6) A induce un cambio de conformación en el grupo metilamino, cambiando de syn a anti, posicionando el grupo metilo en la ranura mayor.
m(6) La A desestabiliza los dúplex de ARN entre 0,5 y 1,7 kcal/mol.
m(6) A estabiliza significativamente las bases de ARN no apareadas en regiones de una sola cadena.
Las adenosinas metiladas muestran una propensión a las estructuras de un solo hilo adyacentes a la modificación.

Conclusiones:

m(6) A exhibe efectos dependientes del contexto sobre la estructura del ARN, desestabilizando los dúplex mientras estabiliza las regiones de un solo hilo.
Estas alteraciones estructurales mediadas por m(6) A probablemente contribuyen a sus funciones reguladoras en el metabolismo del ARN.
Los hallazgos proporcionan información mecanicista sobre cómo m(6)A impacta la función del ARN a nivel estructural.

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