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Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses
Published on: February 26, 2011
Se requiere la proteína de unión al poli (A) para el acortamiento del poli (A) y la iniciación de la traducción
1Department of Biochemistry, Stanford University Medical Center, California 94305.
Cell
|September 8, 1989
Resumen
La proteína de unión poli (PAB) es esencial para el inicio de la traducción en la levadura. Las mutaciones del supresor revelan que la subunidad ribosómica 60S media este requisito de PAB para una traducción eficiente del ARNm.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Genética de la levadura Genética de la levadura
- Síntesis de proteínas Síntesis de proteínas
Sus antecedentes:
- La proteína esencial de unión poli-A (PAB) juega un papel crítico en la regulación del ARNm.
- El agotamiento de PAB en S. cerevisiae inhibe la iniciación de la traducción y acorta las colas poli (A).
Objetivo del estudio:
- Investigar el mecanismo por el cual se requiere PAB para la iniciación de la traducción.
- Para identificar los factores genéticos que pueden suprimir la pérdida de la función PAB.
Principales métodos:
- Utilizando una mutación sensible a la temperatura (pab1-F364L) y la inactivación del promotor para agotar el PAB en S. cerevisiae.
- Realizar análisis de reversión para aislar mutaciones supresoras sensibles al frío (spb1-spb7).
- Analizando los efectos de las mutaciones supresoras en la iniciación de la traducción, la longitud de la cola de poli (A) y la composición de las subunidades ribosómicas.
Principales resultados:
- Se identificaron siete mutaciones extragénicas sensibles al frío (spb1-spb7) que suprimen la deleción de PAB.
- Estas mutaciones permiten la iniciación de la traducción en ausencia de PAB sin alterar las longitudes de la cola de poli (A).
- Se encontró que las mutaciones supresoras afectan a la cantidad de la subunidad ribosómica 60S, con SPB2 codificando la proteína ribosómica L46.
Conclusiones:
- La subunidad ribosómica 60S está implicada en la mediación del requisito de PAB durante la iniciación de la traducción.
- Esta interacción asegura una traducción eficiente del poli(A) + mRNA intacto in vivo.
- Comprender esta vía proporciona información sobre el control de calidad del ARNm y la regulación traslacional.
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Ribosomes translate genetic information encoded by messenger RNA (mRNA) into proteins. Both prokaryotic and eukaryotic cells have ribosomes. Cells that synthesize large quantities of protein—such as secretory cells in the human pancreas—can contain millions of ribosomes.
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Ribosomes are composed of ribosomal RNA (rRNA) and proteins. In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within...
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Ribosomes are composed of ribosomal RNA (rRNA) and proteins. In eukaryotes, rRNA is transcribed from genes in the nucleolus—a part of the nucleus that specializes in ribosome production. Within...
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Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...
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