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Initiation is the first step of transcription in eukaryotes. Prokaryotic RNA Polymerase (RNAP) can bind to the template DNA and start transcribing. On the other hand, transcription in eukaryotes requires additional proteins, called transcription factors, to first bind to the promoter region in the DNA template. This binding helps recruit the specific RNAP that can assemble on the DNA and start transcription.
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RNA polymerase (RNAP) carries out DNA-dependent RNA synthesis in both bacteria and eukaryotes. Bacteria do not have a membrane-bound nucleus. So, transcription and translation occur simultaneously, on the same DNA template.
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Transcription elongation is a dynamic process that alters depending upon the sequence heterogeneity of the DNA being transcribed. Hence, it is not surprising that the elongation complex's composition also varies along the way while transcribing a gene.
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An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication...
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Las estructuras complejas de iniciación de la transcripción elucidan la apertura del ADN

C Plaschka1, M Hantsche1, C Dienemann1

  • 1Max Planck Institute for Biophysical Chemistry, Department of Molecular Biology, Am Fassberg 11, 37077 Göttingen, Germany.

Nature
|May 20, 2016
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores utilizaron la microscopía cryoelectrónica para visualizar los complejos de iniciación de transcripción de levadura. Revelaron cómo el ADN se abre y queda atrapado durante la activación génica, proporcionando un modelo unificado para la iniciación de la transcripción.

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Área de la Ciencia:

  • Biología molecular
  • Biología estructural
  • La genética

Sus antecedentes:

  • La iniciación de la transcripción génica eucariota implica el ensamblaje del complejo de ARN polimerasa (Pol) II y la apertura del ADN promotor.
  • Comprender los mecanismos estructurales precisos de este proceso es crucial para descifrar la regulación génica.

Objetivo del estudio:

  • Determinar las estructuras de alta resolución de los complejos de iniciación de transcripción de levadura en diferentes estados de ADN (cerrado y abierto).
  • Para aclarar las interacciones moleculares que rigen el posicionamiento del ADN, la apertura y la carga de la hebra de la plantilla durante la iniciación de la transcripción.

Principales métodos:

  • Se empleó la criomicroscopia electrónica (crio-EM) para obtener estructuras de complejos de iniciación de levadura.
  • Se obtuvieron datos estructurales de alta resolución (8,8 Å y 3,6 Å) para complejos con ADN cerrado y abierto, respectivamente.

Principales resultados:

  • Las estructuras detalladas revelaron cómo la proteína de unión a la caja TATA (TBP) y los factores de transcripción (TFIIA, TFIIB, TFIIE, TFIIF) posicionan y retienen el ADN dentro de la hendidura Pol II.
  • Se observó una abertura del ADN cerca de la pinza Pol II y el dominio de la "hélice alada extendida" del TFIIE, independiente del TFIIH.
  • La unión alostérica del dominio "E-ribbon" de TFIIE puede facilitar la carga de la hebra de la plantilla al reposicionar los elementos de proteína que lo obstruyen.

Conclusiones:

  • Se propone un modelo unificado para la iniciación de la transcripción, destacando la captura de ADN promotor abierto a través de contactos extensivos proteína-ADN y proteína-proteína.
  • Los hallazgos proporcionan información estructural crítica sobre los primeros eventos de la transcripción génica eucariota.