Temperature-sensitive control of protein activity by conditionally splicing inteins.
Nat Biotechnol
; 22(7): 871-6, 2004 Jul.
Article
en En
| MEDLINE
| ID: mdl-15184905
ABSTRACT
Conditional or temperature-sensitive (TS) alleles represent useful tools with which to investigate gene function. Indeed, much of our understanding of yeast has relied on temperature-sensitive mutations which, when available, also provide important insights into other model systems. However, the rarity of temperature-sensitive alleles and difficulty in identifying them has limited their use. Here we describe a system to generate temperature-sensitive alleles based on conditionally active inteins. We have identified temperature-sensitive splicing variants of the yeast Saccharomyces cerevisiae vacuolar ATPase subunit (VMA) intein inserted within Gal4 and transferred these into Gal80. We show that Gal80-intein(TS) is able to efficiently provide temporal regulation of the Gal4/upstream activation sequence (UAS) system in a temperature-dependent manner in Drosophila melanogaster. Given the minimal host requirements necessary for temperature-sensitive intein splicing, this technique has the potential to allow the generation and use of conditionally active inteins in multiple host proteins and model systems, thereby widening the use of temperature-sensitive alleles for functional protein analysis.
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Bases de datos:
MEDLINE
Asunto principal:
Proteínas Represoras
/
Temperatura
/
Factores de Transcripción
/
Empalme de Proteína
/
Proteínas de Saccharomyces cerevisiae
/
Inteínas
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Límite:
Animals
Idioma:
En
Revista:
Nat Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2004
Tipo del documento:
Article
País de afiliación:
Estados Unidos