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The DNA Replication Fork01:02

The DNA Replication Fork

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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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Replicative Cell Senescence02:15

Replicative Cell Senescence

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Replicative cell senescence is a property of cells that allows them to divide a finite number of times throughout the organism's lifespan while preventing excessive proliferation. Replicative senescence is associated with the gradual loss of the telomere — short, repetitive DNA sequences found at the end of the chromosomes. Telomeres are bound by a group of proteins to form a protective cap on the ends of chromosomes. Embryonic stem cells express telomerase — an enzyme that adds...
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Homologous Recombination02:31

Homologous Recombination

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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...
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DNA Damage can Stall the Cell Cycle02:37

DNA Damage can Stall the Cell Cycle

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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...
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DNA Damage Can Stall the Cell Cycle02:37

DNA Damage Can Stall the Cell Cycle

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Restarting Stalled Replication Forks02:37

Restarting Stalled Replication Forks

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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,...
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Video Experimental Relacionado

Updated: Nov 6, 2025

Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence
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Quantifying Replication Stress in Ovarian Cancer Cells Using Single-Stranded DNA Immunofluorescence

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El estrés de replicación promueve la eliminación celular por extrusión

Vivek K Dwivedi1, Carlos Pardo-Pastor2, Rita Droste1

  • 1Howard Hughes Medical Institute, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.

Nature
|May 6, 2021
PubMed
Resumen

La extrusión celular elimina las células a través del estrés de replicación, un proceso conservado en los animales. Este mecanismo, que involucra a ATR-CHK1 y p53, actúa como un supresor de tumores en los mamíferos.

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Área de la Ciencia:

  • Biología celular
  • Biología del desarrollo
  • La genética

Sus antecedentes:

  • La extrusión celular es un proceso vital de eliminación celular conservado en diversos organismos.
  • La desregulación de la extrusión celular está implicada en enfermedades epiteliales, incluido el cáncer.
  • Los mecanismos moleculares precisos que impulsan la extrusión celular no se comprenden completamente.

Objetivo del estudio:

  • Para investigar los mecanismos subyacentes a la extrusión celular.
  • Identificar los genes que controlan la extrusión celular utilizando una pantalla de todo el genoma en *Caenorhabditis elegans*.
  • Para determinar el papel del estrés de replicación en la extrusión celular.

Principales métodos:

  • Se realizó una prueba de interferencia de ARN de todo el genoma en embriones de Caenorhabditis elegans.
  • Realizó experimentos de imágenes en vivo para analizar la dinámica de la extrusión celular.
  • Se utiliza hidroxiurea para inducir el estrés de replicación en las células epiteliales de mamíferos.

Principales resultados:

  • Se han identificado genes del ciclo celular con funciones específicas de la fase S cruciales para la extrusión.
  • Se ha demostrado que las células de extrusión experimentan y responden al estrés de replicación a través de ATR y CHK1.
  • Se ha demostrado que el bloqueo de la entrada de la fase S o la respuesta al estrés de replicación inhibe la extrusión.
  • Extrusión inducida en células de mamíferos a través del estrés de replicación inducido por la hidroxiurea, dependiente de ATR-CHK1 y p53.

Conclusiones:

  • La extrusión celular inducida por el estrés de replicación se conserva en todas las especies animales.
  • Este proceso es un mecanismo primordial para la eliminación celular.
  • La extrusión celular mediada por el estrés de replicación puede funcionar como supresor tumoral en mamíferos.