A small molecule-directed approach to control protein localization and function.

摘要: Protein localization is tightly linked with function, such that the subcellular distribution of a protein serves as an important control point regulating activity. Exploiting this regulatory mechanism, we present here general approach by which location, and hence may be controlled on demand in budding yeast. In system small molecule, rapamycin, used to temporarily recruit strong cellular address signal target protein, placing under selective chemical stimulus. The kinetics are rapid: rapamycin-directed nucleo-cytoplasmic transport evident 10–12 min post-treatment process reversible upon removal rapamycin. Accordingly, envision platform promising for systematic construction conditional loss-of-function mutants. As proof principle, direct nuclear export essential heat shock transcription factor Hsf1p, thereby mimicking cell-cycle arrest phenotype hsf1 temperature-sensitive mutant. Our drug-induced also provides method can uncoupled from endogenous cell signalling events, addressing necessity or sufficiency given shift particular process. To illustrate, directed import calcineurin-dependent Crz1p absence native stimuli; analysis directly substantiates translocation insufficient its transcriptional total, technology represents powerful generation alleles mislocalization studies yeast, potential applicability genome-wide scale. Copyright © 2008 John Wiley & Sons, Ltd.