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Telomeres and Telomerase02:41

Telomeres and Telomerase

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In eukaryotic DNA replication, a single-stranded DNA fragment remains at the end of a chromosome after the removal of the final primer. This section of DNA cannot be replicated in the same manner as the rest of the strand because there is no 3’ end to which the newly synthesized DNA can attach. This non-replicated fragment results in gradual loss of the chromosomal DNA during each cell duplication. Additionally, it can induce a DNA damage response by enzymes that recognize single-stranded...
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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,...
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Replication in Eukaryotes01:29

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In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
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Translesion DNA Polymerases02:10

Translesion DNA Polymerases

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Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
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Replicative Cell Senescence02:15

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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

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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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In vitro Reconstitution of the Active T. castaneum Telomerase
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La telomerasa se comporta mal después de una ruptura

Nausica Arnoult1, Thomas R Cech2

  • 1Department of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, CO, USA.

Science (New York, N.Y.)
|February 15, 2024
PubMed
Resumen

La supresión de la telomerasa en el ADN roto evita más daños y mantiene la estabilidad del genoma. Este hallazgo es crucial para comprender los mecanismos de reparación del ADN y prevenir la inestabilidad genómica.

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Droplet Digital TRAP ddTRAP: Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction
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Droplet Digital TRAP ddTRAP: Adaptation of the Telomere Repeat Amplification Protocol to Droplet Digital Polymerase Chain Reaction

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Área de la Ciencia:

  • Biología molecular
  • La genética
  • Biología celular

Sus antecedentes:

  • La telomerasa es una enzima clave responsable de mantener la longitud de los telómeros.
  • La actividad incontrolada de la telomerasa puede conducir a la inestabilidad genómica.
  • Las rupturas de ADN son lesiones críticas que deben repararse con precisión.

Objetivo del estudio:

  • Para investigar el papel de la telomerasa en la reparación de la ruptura del ADN.
  • Para determinar si la supresión de la actividad de la telomerasa en las rupturas de ADN afecta la integridad del genoma.

Principales métodos:

  • Utilizó CRISPR-Cas9 para inducir rupturas de doble cadena de ADN dirigidas.
  • El uso de inhibidores de la telomerasa para bloquear la actividad de la enzima en los sitios de ruptura.
  • Se evaluaron los focos de reparación del ADN y las aberraciones cromosómicas mediante análisis microscópicos y moleculares.

Principales resultados:

  • Se observó el reclutamiento de la telomerasa a las rupturas de ADN.
  • La inhibición de la telomerasa en los sitios de ruptura redujo significativamente la reparación aberrante del ADN.
  • La supresión de la actividad de la telomerasa preservó la integridad cromosómica.

Conclusiones:

  • La actividad de la telomerasa en los sitios rotos del ADN puede conducir a resultados perjudiciales.
  • Dirigir la telomerasa a las rupturas del ADN es una estrategia viable para mantener la estabilidad del genoma.
  • Esta investigación ofrece nuevos conocimientos sobre la compleja interacción entre el mantenimiento de los telómeros y la reparación del ADN.