RNA Hairpin Folding in the Crowded Cell. Issue 9 (2nd February 2016)

• Record Type: Journal Article
• Authors: Gao, Mimi; Gnutt, David; Orban, Axel; Appel, Bettina; Righetti, Francesco; Winter, Roland; Narberhaus, Franz; Müller, Sabine; Ebbinghaus, Simon
• Is Part Of: Angewandte Chemie international edition. Volume 55:Issue 9(2016)
• Page Start: 3224
• Page End: 3228
• Publication Date: 2016-02-02
• Subjects: biophysics -- folding stability -- in-cell spectroscopy -- macromolecular crowding -- RNA; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201510847
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: Precise secondary and tertiary structure formation is critically important for the cellular functionality of ribonucleic acids (RNAs). RNA folding studies were mainly conducted in vitro, without the possibility of validating these experiments inside cells. Here, we directly resolve the folding stability of a hairpin‐structured RNA inside live mammalian cells. We find that the stability inside the cell is comparable to that in dilute physiological buffer. On the contrary, the addition of in vitro artificial crowding agents, with the exception of high‐molecular‐weight PEG, leads to a destabilization of the hairpin structure through surface interactions and reduction in water activity. We further show that RNA stability is highly variable within cell populations as well as within subcellular regions of the cytosol and nucleus. We conclude that inside cells the RNA is subject to (localized) stabilizing and destabilizing effects that lead to an on average only marginal modulation compared to diluted buffer. Abstract : From the test tube to the cell : In‐cell folding experiments spatio‐temporally resolve RNA hairpin stability in the living cell. The folding stability inside cells is found to be comparable to that in dilute physiological buffer.