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The Replisome03:01

The Replisome

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DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
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Chromosome Replication

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Before a cell can divide, it must accurately replicate all of its chromosomes, including the DNA and its associated histone and non-histone proteins.  This process begins at numerous origins of replication during the S phase of the cell cycle in each of a cell’s chromosomes simultaneously. Certain nucleotides can act as origins of replication, but these sequences are not well defined - especially in complex, multi-cellular, eukaryotic species. The length of DNA that spans an origin...
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DNA Helicases00:55

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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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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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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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The DNA Replication Fork01:02

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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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Video Experimental Relacionado

Updated: Aug 15, 2025

Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
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Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level

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El complejo de pre-replicación humana es un complejo abierto

Jian Li1, Jiangqing Dong2, Weitao Wang3

  • 1School of Biological Sciences, The University of Hong Kong, Hong Kong, China.

Cell
|January 7, 2023
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores descubrieron cómo el mantenimiento del minicromosoma (MCM) el hexámero doble (DH) inicia la replicación del ADN mediante la fusión de hebras de ADN. Esta estructura revela una estructura abierta inicial (IOS) crucial para la licencia de replicación en células humanas.

Palabras clave:
Iniciación de la replicación del ADNcomplejo MCM27 humanofusión inicial del ADNFuego de origenPrevio a la RCLicencias de replicación

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Área de la Ciencia:

  • Biología molecular
  • La genética
  • La bioquímica

Sus antecedentes:

  • El inicio de la replicación del ADN es esencial para la división celular en los eucariotas.
  • El mantenimiento del minicromosoma (MCM) 2-7 doble hexámero (DH) es un complejo clave para iniciar la replicación del ADN.
  • El mecanismo preciso por el cual el MCM-DH funde el ADN de origen sigue siendo en gran medida desconocido.

Objetivo del estudio:

  • Aclarar el mecanismo estructural de la fusión inicial del ADN por el MCM-DH humano (hMCM-DH) durante el inicio de la replicación.
  • Comprender cómo el hMCM-DH facilita la formación de una estructura abierta inicial (IOS) en los orígenes de la replicación.
  • Para investigar el papel del IOS en la licencia de la replicación del ADN.

Principales métodos:

  • Microscopía crioelectrónica (cryo-EM) para determinar la estructura de alta resolución de la hMCM-DH unida al ADN.
  • Ensayos bioquímicos para evaluar el impacto de la alteración de la SIO en la formación de DH y el inicio de la replicación.
  • Huella genómica para mapear la distribución de hMCM-DH y IOS en todo el genoma.

Principales resultados:

  • Una estructura cryo-EM de 2.59-Å de la hMCM-DH reveló un canal central estrecho que distorsiona el ADN unido.
  • El hMCM-DH induce una estructura abierta inicial (IOS) al separar un par de bases dentro del dúplex de ADN distorsionado.
  • La alteración de la IOS inhibe significativamente la formación de MCM-DH y el posterior inicio de la replicación.
  • El mapeo genómico mostró que las IOS se agrupan en zonas de iniciación, en correlación con el disparo de origen estocástico.

Conclusiones:

  • El hMCM-DH posee un mecanismo intrínseco para acoplar la unión del ADN con la separación inicial de la cadena, formando un IOS.
  • Este IOS es crítico para la licencia de inicio de la replicación del ADN en células humanas.
  • Los hallazgos proporcionan una base estructural para comprender cómo se seleccionan y abren los orígenes de replicación.