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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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Photoactivated Localization Microscopy with Bimolecular Fluorescence Complementation (BiFC-PALM)

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Coconversión de secuencias de flanqueo con conmutación homotálica.

C McGill¹, B Shafer, J Strathern

¹Laboratory of Eukaryotic Gene Expression, National Cancer Institute, Frederick Cancer Research Facility, Maryland 21701.

|May 5, 1989

Resumen

La conmutación homotálica en S. cerevisiae reemplaza el ADN del tipo de apareamiento. Se observaron gradientes variables de extensión de reemplazo y de coconversión, lo que sugiere un intermediario de heterodúplex de ADN durante el cambio de tipo de apareamiento.

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Área de la Ciencia:

Biología Molecular Biología Molecular
Genética La genética.
Biología de la levadura Biología de la levadura.

Sus antecedentes:

La conmutación homotálica en Saccharomyces cerevisiae es un proceso de conversión genética de tipo pareado.
El locus de tipo de apareamiento (MAT) intercambia ADN con los locus silenciosos (HML o HMR).

Objetivo del estudio:

Para investigar la variabilidad en la extensión de reemplazo de ADN durante la conmutación homotálica.
Para analizar el gradiente de coconversión a través de la región X del locus MAT.
Para explorar las implicaciones de las diferencias de sitio de flanqueo para los mecanismos de reparación del ADN.

Principales métodos:

Utilizando polimorfismos del sitio de restricción para diferenciar entre MAT y los loci del donante.
Analizando los patrones de reemplazo de secuencias de ADN durante el cambio de tipo de apareamiento.
Examinando los eventos de coconversión en relación con la escisión de doble hebra mediada por el producto genético HO.

Principales resultados:

Demostrado grado variable de reemplazo de ADN MAT durante la conmutación homotálica.
Se identificó un gradiente de coconversión a través de la región X.
Se observó coconversión en ambos lados de la ruptura de doble hebra.
Se notaron diferentes sitios de flanqueo en las células hijas, indicativos de un intermediario heterodúplex.

Conclusiones:

La extensión de la sustitución del ADN durante el cambio de tipo de apareamiento de S. cerevisiae no es fija.
Es probable que un intermediario heterodúplex de ADN esté involucrado en el proceso de coconversión.
La comprensión de estos mecanismos proporciona información sobre la conversión de genes y las vías de reparación del ADN.