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IX - Mayer/Möglich

A Bacterial Light-Activated RNA-Binding Protein as an Orthogonal Optogenetic Actuator in Eukaryotes

Participants:

1) Günter Mayer (PI; University of Bonn), coworker: Anna Pyka

2) Andreas Möglich (PI; University of Bayreuth), coworker:

Abstract:

A Bacterial Light-Activated RNA-Binding Protein as an Orthogonal Optogenetic Actuator in Eukaryotes

As genetically encodable, light-regulated actuators, sensory photoreceptors underpin optogenetics. Whereas the pioneering optogenetic studies exclusively relied on certain rhodopsin photoreceptors that serve as light-gated cation channels or lightdriven ion pumps, numerous other photoreceptors have since been discovered in Nature or have been engineered that now allow the spatiotemporally precise, reversible and non-invasive control by light of manifold cellular processes and properties. Although several robust photoreceptor systems are in place, which afford optogenetic intervention at the DNA and protein levels, there is a notable lack of systems that would directly act at the RNA level. To fill this gap, we will establish a bacterial, blue-light-activated RNA-binding photoreceptor, denoted PAL, as an orthogonal optogenetic instrument in eukaryotic cells in order to allow the deliberate perturbation and control of cellular events in unprecedented ways. A thorough biochemical and biophysical characterization will unravel the signal transduction mechanisms employed by PAL and will inform the engineering of derivative and improved light-regulated RNA-binding proteins. In vitro evolution will be employed to optimize and diversify the RNA targets PAL binds to. Thus empowered, we will establish in eukaryotic cell culture PAL-based systems for regulation by light of the translation of arbitrary target proteins. Robust, light-regulated systems will be transferred to the animal model Drosophila melanogaster. We note that light-regulated binding of RNA represents a new optogenetic modality that not only applies to translation but to numerous other RNA-associated processes. We are thus confident that PAL will become an important, widely applicable and previously unavailable optogenetic actuator.

 

Publications Mayer:

Selective aptamer-based control of intraneuronal signaling. Lennarz S., Alich T.C., Kelly T., Blind M., Beck H., und Mayer G. Angew. Chem. Int. Ed. 2015, 54, 5369-5373.

An aptamer to the MAP kinase insert region. Lennarz S., Heider E., Blind M., und Mayer G. ACS Chem. Biol., 2015, 10, 320-327.

Aptamer-guided caging for selective masking of protein domains. Rohrbach F., Schäfer F.,Fichte M.A.H., Pfeiffer F., Mu?ller J., Pötzsch B., Heckel A., und Mayer G. Angew. Chem. Int. Ed. 2013, 52, 1912-1915.

Molecular mechanism for inhibition of G protein-coupled receptor kinase 2 by a selective RNA aptamer. Tesmer V.M., Lennarz S., Mayer G., und Tesmer J.J. Structure 2012, 20, 1300-1309.

Carba-sugars activate the glmS-riboswitch from Staphylococcus aureus. Lu?nse C.E., Schmidt M., Wittmann V., und Mayer G. ACS Chem Biol., 2011, 6, 675-678.