title: 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.
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohtsuki
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Shigeto
authors.initials: S
authors.lastname: Kanzaki
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Sae
authors.initials: S
authors.lastname: Nishimura
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Kunihiro
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Masahiko
authors.initials: M
authors.lastname: Sisido
authors.affiliation: Department of Biomedical Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Watanabe
mesh_terms: D000215:Acylation; D000818:Animals; D016466:CHO Cells; D003062:Codon / Q000235:genetics; D003374:Coumarins / Q000737:chemistry*; D006224:Cricetinae; D003412:Cricetulus; D016368:Frameshift Mutation / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007834:Lasers; D008027:Light; D008081:Liposomes / Q000737:chemistry; D010444:Peptide Elongation Factor Tu / Q000378:metabolism; D010782:Photolysis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*
chemical_list: D003062:Codon; D003374:Coumarins; D008081:Liposomes; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C030123:coumarin; D010444:Peptide Elongation Factor Tu