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

Takashi Ohtsuki (), Shigeto Kanzaki, Sae Nishimura, Yoshio Kunihiro, Masahiko Sisido and Kazunori Watanabe
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Takashi Ohtsuki: Okayama University
Shigeto Kanzaki: Okayama University
Sae Nishimura: Okayama University
Yoshio Kunihiro: Okayama University
Masahiko Sisido: Okayama University
Kazunori Watanabe: Okayama University

Nature Communications, 2016, vol. 7, issue 1, 1-7

Abstract: 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.

Date: 2016
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DOI: 10.1038/ncomms12501

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