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Recuperación de la actividad mitogénica de un mutante del factor de crecimiento con una secuencia de translocación
1Laboratory of Molecular Biology, Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross, Rockville, MD 20855.
Science (New York, N.Y.)
|September 28, 1990
Resumen
La translocación nuclear es crucial para el factor de crecimiento-1 de unión a la heparina.
Área de la Ciencia:
- Biología Molecular Biología Molecular
- Biología celular Biología celular.
- La bioquímica es la bioquímica.
Sus antecedentes:
- El factor de crecimiento de unión a la heparina-1 (HBGF-1) es un polipéptido angiogénico mitógeno.
- El HBGF-1 es biológicamente activo después del truncamiento amino-terminal (HBGF-1 alfa).
Objetivo del estudio:
- Investigar el papel de una supuesta secuencia de translocación nuclear en la actividad mitogénica del HBGF-1.
- Para determinar si se requiere la localización nuclear para la proliferación celular inducida por HBGF-1.
Principales métodos:
- La mutagenesis de la reacción en cadena de la polimerasa para crear HBGF-1U, carece de la secuencia de translocación nuclear.
- Sistemas de expresión procariótica para producir proteínas HBGF-1 mutantes.
- Ensayos para la síntesis de ADN, la proliferación celular, la fosforilación de tirosina y la expresión de c-fos.
Principales resultados:
- HBGF-1U conservó la capacidad de unión a la heparina, pero no pudo inducir la síntesis de ADN y la proliferación celular.
- HBGF-1U indujo la fosforilación de tirosina mediada por el receptor intracelular y la expresión de c-fos.
- Un polipéptido quimérico (HBGF-1U2) con una secuencia de translocación nuclear recuperó la actividad mitogénica.
Conclusiones:
- La translocación nuclear es esencial para la actividad mitogénica de HBGF-1.
- La secuencia de translocación nuclear media la respuesta biológica del HBGF-1 en la proliferación celular.
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