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Denise Raddatz Mota
Genome-Wide Analysis of the Core DNA Replication Machinery in the Higher Plants Arabidopsis and Rice
Randall W. Shultz2, Vinaya M. Tatineni, Linda HanleyBowdoin*, and William F. Thompson
Arroz (Oryza sativa)
Arabidopsis thaliana
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Replicación del ADN en Eucariontes
• Numerosos complejos de multiproteínas.- Iniciación- Elongación- Maduración de los fragmentos de
Okasaki
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¿Qué se sabe sobre la Maquinaria de la Replicación del ADN?
Levaduras
Modelo Animal Plantas
• Combinación de información experimental publicada con análisis bioinformáticos de la secuencia genómica en Arabidopsis y arroz.
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Estrategia• Ensamblaje de base de datos a partir de las proteínas
(maquinaria de replicación del ADN) de levadura y animal.
• Se obtuvo la base de datos del genoma traducido de Arabidopsis con el algoritmo BLAST.
• Las secuencias de Arabidopsis se utilizaron para encontrar homólogos en arroz.
• Para el análisis filogenético de las proteínas se utilizó el programa ClustalW.
• Para el alineamiento de las múltiples secuencias de complejos de proteína se utilizó el algoritmo ClustalW.
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Maquinaria de la Replicación
del ADN en Eucariontes
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Pre-iniciación
Complejo de pre-replicación (pre-RC): ORC1-6 + CDC6 + MCM2-7
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Iniciación
Supercomplejo con función de helicasa: GINS + MCM2-7 +CDC45
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Elongación
GINS + MCM2-7 +CDC4 5 + Pol E
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Maduración
Fragmentos de Okasaki:Pol D + FEN1 + LIG 1 + PCNA
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Genes de la Maquinaria de Replicación de ADN en la Pre-Iniciación
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Genes de la Maquinaria de Replicación de ADNen la Iniciación
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Genes de la Maquinaria de Replicación de ADNen la Elongación
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Genes de la Maquinaria de Replicación de ADNen la Elongación
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Genes de la Maquinaria de Replicación de ADN en la Maduración
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Alineamientos de Secuencias Múltiples de proteína ORC6
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Alineamientos de Secuencias Múltiples de proteína PSF1
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Alineamientos de Secuencias Múltiples de proteína PSF2
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Alineamientos de Secuencias Múltiples de proteína PSF3
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Alineamientos de Secuencias Múltiples de proteína SLD5
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PSF1 PSF2 PSF3
SLD5
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Múltiples Copias de Genes de la Maquinaria de Replicación de ADN
GENE Humano Levadura Arabidopsis ArrozORC1 1 1 2 1CDC6 1 1 2 1CDT1 1 1 2 2CDC45 1 1 1 2TOPBP1 1 1 2 3PSF3 1 1 2 1POLD4 1 0 1 2POLE1 1 1 2 1POLE2 1 1 1 2POLE3 1 1 GF GFPOLE4 1 1 GF GFPCNA 1 1 2 1RPA1 1 1 5 3RPA2 2 1 2 3RPA3 1 1 2 1FEN1 1 1 1 2LIG1 1 1 3 1
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Conclusiones• La maquinaria de replicación del ADN de
animales y levaduras, está conservada en plantas, mostrando muchas características en común con otros eucariontes y otras que son únicas en plantas.
• Las proteínas de replicación del ADN en plantas, son más similares a las del humano que a las de levaduras.
• Los modelos animales pueden ser más importantes para el estudio de replicación del ADN en plantas que los de levadura.