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DNA Topoisomerases02:02

DNA Topoisomerases

Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
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Topoisomerases are divided into two main types.  Type I...
DNA Damage can Stall the Cell Cycle02:36

DNA Damage can Stall the Cell Cycle

In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
Homologous Recombination02:31

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
Restarting Stalled Replication Forks02:37

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DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
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Published on: March 25, 2020

Las cinasas dependientes de la ciclina previenen la replicación del ADN a través de múltiples mecanismos.

V Q Nguyen1, C Co, J J Li

  • 1Department of Biochemistry and Biophysics, University of California, San Francisco 94143-0414, USA.

Nature
|June 29, 2001
PubMed
Resumen

La prevención de la reiniciación de la replicación del ADN es crucial para el control del ciclo celular. En la levadura, las cinasas dependientes de ciclina (CDK) de tipo B utilizan tres mecanismos para bloquear la replicación, asegurando la estabilidad del genoma.

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Área de la Ciencia:

  • Biología celular Biología celular.
  • Biología Molecular Biología Molecular
  • Genética La genética.

Sus antecedentes:

  • La replicación fiel del genoma una vez por ciclo celular es esencial para la estabilidad genética.
  • La replicación del ADN eucariota se inicia en múltiples orígenes, lo que debe evitarse que se reinicie dentro del mismo ciclo celular.
  • Se sabe que las cinasas dependientes de ciclina (CDK) juegan un papel en el bloqueo de la reiniciación de la replicación, pero los mecanismos precisos siguen siendo elusivos.

Objetivo del estudio:

  • Para dilucidar los mecanismos por los cuales las cinasas dependientes de ciclina tipo B (CDKs) impiden la reiniciación de la replicación del ADN en Saccharomyces cerevisiae.
  • Determinar la importancia funcional de las vías inhibidoras individuales reguladas por las CDK.

Principales métodos:

  • Investigó el papel de las CDK de tipo B en la prevención de la reiniciación de la replicación en la levadura.
  • Se analizaron los efectos de la actividad de CDK en la fosforilación del complejo de reconocimiento de origen (ORC), la actividad de Cdc6 y la localización del complejo Mcm2-7.
  • Se evaluaron las consecuencias de interrumpir estas vías reguladoras en la replicación del ADN en las células de fase G2/M.

Principales resultados:

  • Las CDK de tipo B en Saccharomyces cerevisiae emplean múltiples mecanismos superpuestos para inhibir la reiniciación de la replicación.
  • Estos mecanismos incluyen la fosforilación del complejo de reconocimiento de origen (ORC), la regulación a la baja de la actividad de Cdc6 y la exclusión nuclear del complejo Mcm2-7.
  • Solo cuando las tres vías inhibidoras se interrumpen simultáneamente, los orígenes de la replicación reinician la replicación del ADN en las células G2/M.

Conclusiones:

  • Las CDK de tipo B utilizan una estrategia inhibidora multifacética para asegurar una sola ronda de replicación del ADN por ciclo celular.
  • Cada uno de los mecanismos reguladores identificados (fosforilación de ORC, regulación a la baja de Cdc6, exclusión de Mcm2-7) es funcionalmente significativo para prevenir la reiniciación.
  • Estos hallazgos proporcionan una comprensión completa de cómo las células eucariotas mantienen la estabilidad del genoma mediante el control estricto de la replicación del ADN.