Phototriggered protein syntheses by using (7-diethylaminocoumarin-4-yl)methoxycarbonyl-caged aminoacyl tRNAs.

ABSTRACT: The possibility of spatiotemporally photocontrolling translation holds considerable promise for studies on the biological roles of local translation in cells and tissues. Here we report caged aminoacyl-tRNAs (aa-tRNAs) synthesized using a (7-diethylaminocoumarin-4-yl)methoxycarbonyl (DEACM)-cage compound. DEACM-caged aa-tRNA does not spontaneously deacylate for at least 4?h in neutral aqueous solution, and does not bind to the elongation factor Tu. On irradiation at ?405?nm at 125?mW?cm(-2), DEACM-aa-tRNA is converted into active aa-tRNA with a half-life of 19?s. Notably, this rapid uncaging induced by visible light does not impair the translation system. Translation is photoinduced when DEACM-aa-tRNA carrying a CCCG or a CUA anticodon is uncaged in the presence of mRNAs harbouring a CGGG four-base codon or a UAG amber codon, respectively. Protein synthesis is phototriggered in several model systems, including an in vitro translation system, an agarose gel, in liposomes and in mammalian cells.

SUBMITTER: Ohtsuki T 

PROVIDER: S-EPMC4992060 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature