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Formación de un dímero cuadruplex entrelazado por d(GGGT)

Yamuna Krishnan-Ghosh¹, Dongsheng Liu, Shankar Balasubramanian

¹Contribution from the University Chemical Laboratories, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K.

Journal of the American Chemical Society

|September 2, 2004

Resumen

El tetranucleótido d ((GGGT) se autoensambla en dímeros cuadruplexos paralelos. Se identificaron dos especies distintas, un tetrámero y un octámero, revelando nuevos arreglos estructurales que incluyen un dímero cuadruplex deslizado con una única tetra GTGT.

Área de la Ciencia:

La bioquímica es la bioquímica.
Biología Molecular Biología Molecular
Biología Estructural Biología estructural.

Sus antecedentes:

Las secuencias de ADN pueden autoensamblarse en estructuras complejas.
Los G-cuadruplexos son estructuras de ADN no canónicas con diversas funciones biológicas.
Se sabe que la secuencia d ((GGGT) forma estructuras cuadruplexas.

Objetivo del estudio:

Para investigar el autoensamblaje del tetranucleótido d(GGGT).
Para caracterizar las estructuras de las especies cuadruplex resultantes.
Para dilucidar el mecanismo de la formación de dímeros cuádruples.

Principales métodos:

Estudios de fusión UV para identificar especies distintas y su estabilidad térmica.
Espectroscopia de dicroísmo circular (CD) para determinar la topología cuadruplex.
Espectrometría de masas de ionización nano-electrospray de iones negativos (nano-ESI-MS) para determinar la estequiometría compleja.
Espectroscopia de Resonancia Magnética Nuclear (RMN) (1) H de alta resolución para resolver la estructura a nivel atómico.

Principales resultados:

Se identificaron dos especies cuádruples distintas, Q (tetrámeras, d (GGGT) y Q (h) (octámeras, d (GGGT) y 8)).
Ambas especies exhibieron espectros de CD característicos de los cuádruplexos paralelos.
La RMN de alta resolución confirmó d{\displaystyle d} como un cuadruplex tetramolecular paralelo C{\displaystyle C} simétrico.
La estructura d ((GGGT) ((8) implica la dimerización de un cuadruplejo paralelo "deslizado", formando un tetrado G adicional y un nuevo tetrado GTGT en los extremos 3'.

Conclusiones:

La secuencia d ((GGGT) forma cuadruplexos paralelos estables y sus dímeros.
Se descubrió una nueva arquitectura de dímeros cuadruplex "deslizada", que involucra la formación de tetragrama G intermolecular.
El estudio revela nuevos conocimientos sobre la diversidad estructural del G-cuadruplex y los mecanismos de autoensamblaje.

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