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Regulation of mRNA translation by a photoriboswitch

KA. Rotstan, MM. Abdelsayed, LF. Passalacqua, F. Chizzolini, K. Sudarshan, AR. Chamberlin, J. Míšek, A. Luptak

. 2020 ; 9 (-) : . [pub] 20200213

Language English Country Great Britain

Document type Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.

Persistent link   https://www.medvik.cz/link/bmc21020742

Grant support
1804220 National Science Foundation
99999.013571/2013-03 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
17-25897Y Grantová Agentura České Republiky
R01 GM094929 NIGMS NIH HHS - United States
T32 GM108561 NIGMS NIH HHS - United States
17-25897Y Grantová Agentura ?eské Republiky

Optogenetic tools have revolutionized the study of receptor-mediated processes, but such tools are lacking for RNA-controlled systems. In particular, light-activated regulatory RNAs are needed for spatiotemporal control of gene expression. To fill this gap, we used in vitro selection to isolate a novel riboswitch that selectively binds the trans isoform of a stiff-stilbene (amino-tSS)-a rapidly and reversibly photoisomerizing small molecule. Structural probing revealed that the RNA binds amino-tSS about 100-times stronger than the cis photoisoform (amino-cSS). In vitro and in vivo functional analysis showed that the riboswitch, termed Werewolf-1 (Were-1), inhibits translation of a downstream open reading frame when bound to amino-tSS. Photoisomerization of the ligand with a sub-millisecond pulse of light induced the protein expression. In contrast, amino-cSS supported protein expression, which was inhibited upon photoisomerization to amino-tSS. Reversible photoregulation of gene expression using a genetically encoded RNA will likely facilitate high-resolution spatiotemporal analysis of complex RNA processes.

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