Análisis genéticos de la maquinaria de transcripción en la levadura Saccharomyces cerevisiae
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2016-03-14
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Jaén: Universidad de Jaén
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[ES] La transcripción en levaduras, como en el resto de eucariotas, es llevada a cabo por las ARN polimerasas I, II y III y en su biogénesis parecen intervenir unos complejos de prefoldinas. Asociada a estos complejos se encuentra la proteína Bud27 en Saccharomyces cerevisiae, ortóloga a la proteína URI/RMP en humanos. Bud27 también media la interacción de la ARN pol. II con los genes y la elongación de la transcripción. En este trabajo de investigación, se ha utilizado una cepa de S.cerevisiae bud27Δ, con deleción para el gen BUD27, cuya mutación da lugar a un fenotipo de termosensibilidad. Se transformó con una genoteca de S.cerevisiae en plásmido multicopia, para buscar los supresores de dicho fenotipo. Tras la realización de diferentes escrutinios, se obtuvo un total de 93 supresores, con alta eficiencia. La posterior identificación de los genes supresores, permitirá relacionarlos con la proteína Bud27 y su función en la transcripción.
[EN] The transcription in yeast, as in other eukaryote organisms, is carried out by RNA polymerases I, II and III and prefoldin complexes seem to take part in their biogenesis. Bud27 protein, whose orthologue in humans is the unconventional prefoldin URI/RMP, is associated to these complexes and is demonstrated to be the first protein involved in assembly of the three RNA pols. Bud27 also mediates RNA pol. II interaction with the genes and the transcription elongation. In this research work, a S.cerevisiae bud27Δ mutant strain, containing the deletion of the BUD27 gene, has been used, whose mutation originates a thermo-sensitive phenotype. This strain was transformed with a library containing the S. cerevisiae genome in multicopy plasmid to search for suppressors of this phenotype. Once the screening performed, we obtain 93 suppressors, with high efficiency. The identification of suppressor genes will allow to analyze the association of these genes with Bud27 protein and its function in transcription.
[EN] The transcription in yeast, as in other eukaryote organisms, is carried out by RNA polymerases I, II and III and prefoldin complexes seem to take part in their biogenesis. Bud27 protein, whose orthologue in humans is the unconventional prefoldin URI/RMP, is associated to these complexes and is demonstrated to be the first protein involved in assembly of the three RNA pols. Bud27 also mediates RNA pol. II interaction with the genes and the transcription elongation. In this research work, a S.cerevisiae bud27Δ mutant strain, containing the deletion of the BUD27 gene, has been used, whose mutation originates a thermo-sensitive phenotype. This strain was transformed with a library containing the S. cerevisiae genome in multicopy plasmid to search for suppressors of this phenotype. Once the screening performed, we obtain 93 suppressors, with high efficiency. The identification of suppressor genes will allow to analyze the association of these genes with Bud27 protein and its function in transcription.
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Genética