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The basic structure of RNA consists of a string of ribonucleotides attached by phosphodiester bonds. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in regulating gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
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Exponiendo los estados de conformación de ARN de alta afinidad ocultos

Nicole I Orlovsky1, Hashim M Al-Hashimi1,2, Terrence G Oas1,2

  • 1Department of Biochemistry , Duke University Medical Center , Durham , North Carolina 27710 , United States.

Journal of the American Chemical Society
|December 10, 2019
PubMed
Resumen
Este resumen es generado por máquina.

La comprensión de la afinidad de unión al ARN requiere tener en cuenta las sanciones conformacionales. Este estudio revela que incluso la unión general débil de ARN-ligando puede implicar una fuerte unión a estados específicos de ARN pre-doblado, lo que afecta el descubrimiento de fármacos.

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Área de la Ciencia:

  • La bioquímica
  • Biología estructural
  • Biofísica molecular

Sus antecedentes:

  • La afinidad de unión al ARN está influenciada por los cambios conformacionales y los costos de energía asociados, denominados "penalidades conformacionales".
  • La medición de estas sanciones es un desafío ya que las conformaciones de ARN unidas a menudo tienen poblaciones indetectables en el estado no unido.

Objetivo del estudio:

  • Desarrollar un modelo termodinámico para el cambio conformacional de ARN acoplado y la unión de ligandos.
  • Cuantificar las sanciones conformacionales en el sistema de interacción ARN-argininamida (ARG) del TAR del VIH-1.

Principales métodos:

  • Se utilizan las perturbaciones de desplazamiento químico y la dispersión de relajación por resonancia magnética nuclear (RMN).
  • Aplicó la inferencia bayesiana a un modelo de 12 estados para estimar las energías de los estados de baja población.
  • Se investigó la interacción entre el ARN TAR del VIH-1 y la argininamida (ARG) como un imitador de la proteína Tat.

Principales resultados:

  • Se identificaron al menos cuatro ARN intermedios distintos basados en la conformación TAR y la ocupación ARG.
  • Se reveló que la conformación de TAR vinculada dominante, con dos ligandos ARG, tiene una población de equilibrio por debajo del límite de detección no vinculada.
  • Se ha demostrado que el ARG se une a la conformación de TAR pre-doblada con afinidad nanomolar, a pesar de una débil afinidad general aparente (0,2 mM).

Conclusiones:

  • Las sanciones conformacionales determinan significativamente la afinidad de unión de ARN-ligando y pueden impulsar la cooperatividad de unión.
  • La predicción precisa del reconocimiento de ARN requiere considerar los costos de energía de las transiciones conformacionales.
  • Los hallazgos tienen implicaciones para el descubrimiento de fármacos dirigidos al ARN y la comprensión de las interacciones entre ARN y proteínas.