GAL4 is a substrate for caspases: implications for two-hybrid screening and other GAL4-based assays.
Mol Cell Biol Res Commun
; 1(2): 158-61, 1999 May.
Article
em En
| MEDLINE
| ID: mdl-10356366
ABSTRACT
Yeast two-hybrid technology as well as mammalian reporter assays use fusions between a protein of interest and the GAL4 DNA-binding domain (GAL4DB). We demonstrate that expression of a GAL4DB/caspase-1 chimeric protein in yeast leads to autoproteolytic cleavage of GAL4DB. Moreover, recombinant GAL4DB is a good in vitro substrate for recombinant caspase-1 and several other caspases. Cleavage sites map at the C-terminus of GAL4DB and result in release of the fused protein. The finding that GAL4DB can be cleaved by caspases has important implications for the use of caspases in two-hybrid analysis and in the interpretation of mammalian assays based on GAL4-dependent reporter gene expression.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fatores de Transcrição
/
Proteínas Recombinantes de Fusão
/
Proteínas Fúngicas
/
Caspase 1
/
Proteínas de Saccharomyces cerevisiae
/
Proteínas de Ligação a DNA
Tipo de estudo:
Diagnostic_studies
/
Screening_studies
Idioma:
En
Revista:
Mol Cell Biol Res Commun
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
1999
Tipo de documento:
Article
País de afiliação:
Bélgica