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Clonado y estructura de dominio del factor de empalme de mamíferos U2AFAF
P D Zamore1, J G Patton, M R Green
1Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Nature
|February 13, 1992
Resumen
Los investigadores aislaron y expresaron un clon de ADN complementario para la subunidad grande del factor auxiliar U2 snRNP 65 (U2AF65). Este factor de empalme esencial contiene distintos dominios de unión al ARN y específicos para el empalme.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Mecanismos de empalme del ARN Mecanismos de empalme del ARN
- Regulación de la expresión génica Regulación de la expresión génica
Sus antecedentes:
- El empalme del ARN pre-mensajero (ARNm) es un proceso crítico en la expresión génica.
- El factor auxiliar U2 snRNP (U2AF) es esencial para el empalme del ARNm de los mamíferos.
- U2AF comprende una subunidad grande (U2AF65) y una pequeña subunidad.
Objetivo del estudio:
- Para aislar y expresar el ADN complementario (ADNc) que codifica la subunidad grande de U2AF (U2AF65).
- Para caracterizar los dominios funcionales de la proteína U2AF65.
- Para comprender la base molecular del papel de U2AF65 en el empalme del pre-ARNm.
Principales métodos:
- La clonación y el aislamiento del ADN complementario (ADNc).
- La expresión in vitro de la proteína U2AF65.
- Análisis del dominio funcional de U2AF65.
Principales resultados:
- Un clon de ADNc que codifica la subunidad grande U2AF65 fue aislado con éxito y expresado in vitro.
- La proteína U2AF65 contiene dos dominios funcionales distintos.
- Un dominio exhibe la unión de ARN específica de la secuencia a través de los dominios de la secuencia de consenso de ribonucleoproteína (RNP-CS).
- El otro dominio presenta un motivo rico en arginina / serina crucial para el empalme, pero no la unión directa al pre-ARNm.
Conclusiones:
- El ADNc aislado U2AF65 codifica una proteína funcional con dominios caracterizados.
- Los dominios RNP-CS median la unión de ARN específica de la secuencia.
- El motivo rico en arginina/serina es esencial para la actividad de empalme de U2AF65.
- Estos hallazgos proporcionan información sobre los mecanismos moleculares de U2AF65 en el empalme de pre-ARNm.
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