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Identificación de una nueva estructura de ramificación Y como intermedio en el procesamiento del ARNm tripanosómico:

W J Murphy, K P Watkins, N Agabian

|November 21, 1986

Resumen

Los ARN mensajeros del tripanosoma (ARNm) adquieren sus extremos 5' a través del empalme del ARN trans. Este proceso implica un ARN líder empalmado (SL) que dona su secuencia al ARNm.

Área de la Ciencia:

Biología Molecular Biología Molecular
Genética La genética.
ARN Biología Biología ARN

Sus antecedentes:

Los ARNm de Trypanosoma brucei poseen una secuencia única de líder empalmado (SL) de 5'.
El mecanismo de adición de la secuencia SL a los ARNm de T. brucei no estaba claro anteriormente.

Objetivo del estudio:

Proporcionar evidencia para el empalme de ARN trans como el mecanismo para la adición de secuencias SL.
Caracterizar las estructuras intermedias involucradas en este proceso.

Principales métodos:

Análisis de la nucleasa S1 para detectar fragmentos de ARN.
La hibridación del norte para evaluar las especies de ARN.
Tratamiento con enzimas de desbranqueamiento para liberar fragmentos de ARN.

Principales resultados:

Se detectó un fragmento de 100 nucleótidos (100-mer) del ARN SL.
El 100-mer está vinculado al ARN poli(A) + a través de un enlace de 2 ransformers-5 'fosfodiéster.
La enzima de desbranqueamiento liberó el 100-mer, lo que indica un intermedio ramificado.

Conclusiones:

El ARN líder empalmado (SL) es el donante de la secuencia SL a los ARNm de T. brucei.
El empalme de ARN trans, no el empalme cis, es el mecanismo para la adición de SL.
Un intermedio ramificado con una estructura en Y confirma el modelo de empalme trans.

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