The long term goal is to be able to correctly predict from its sequence (1) how an RNA folds into its biologically functional form, (2) how its structure affects its function, and (3) how fast it folds and responds to proteins and other ligands in its environment. RNA molecules are crucial in all aspects of gene expression. Messenger RNAs transfer the genetic information from DNA to proteins;in RNA viruses the RNA is both the genetic information, and the messenger for protein synthesis. Non-coding RNAs, including microRNAs and small interfering RNAs, regulate transcription of DNA to RNA and translation of RNA to protein. Errors in processing and control of messenger RNAs are linked to human diseases, including neurodegenerative diseases such as Lou Gehrig's disease. RNAs are being investigated as inhibitors of messenger RNAs to silence specific genes, and to cure diseases. RNAs - in humans, viruses, and bacteria - are also outstanding targets for drugs to prevent or cure human diseases. Knowledge of the three-dimensional structures of RNAs, their stabilities, and their rates of interconversion is crucial to understanding all the different biological functions of RNA. In order to obtain this knowledge, newly discovered single-molecule methods are applied: Laser Tweezers and Fluorescence Resonance Energy Transfer (FRET). RNA molecules are synthesized in the laboratory by RNA polymerase from a DNA template. A specific label is attached to each RNA molecule so that it can be monitored and characterized. Laser Tweezers: A micron-sized bead is attached to each end of a single RNA molecule;one bead is held in a micropipette, the other in a laser trap. As the RNA molecule moves, or changes shape as it interacts with other RNAs or proteins, the beads move. Their motion can be seen in a microscope and thus the behavior of the RNA is assessed. FRET: Fluorescent dyes are placed on the RNA or on the molecules it interacts with. The fluorescent signals from the dyes are exquisitely sensitive to the distance between the dyes, and thus on the motion and shape of the RNA. These data provide the thermodynamic stability of the RNA, and its rates of unfolding and refolding. The effects of proteins that unwind, transcribe, and translate the RNA, as well as drugs that inhibit these processes are determined. This information will lead to improved understanding of RNA structure, stability, and dynamics. It will help in understanding RNA function, and in controlling the role of RNA in human diseases.
RNA viruses cause many human diseases including AIDS, flu, hepatitis C, SARS, etc. Malfunctioning RNAs that occur in humans can also cause diseases, such as Parkinson's disease. Investigation of RNA structure will provide better understanding of RNA function, and can lead to better drugs to cure or prevent human disease.
| Goldman, Daniel H; Kaiser, Christian M; Milin, Anthony et al. (2015) Ribosome. Mechanical force releases nascent chain-mediated ribosome arrest in vitro and in vivo. Science 348:457-60 |
| Tinoco Jr, Ignacio (2015) The ethical scientist: An old-fashioned view. Biopolymers 103:424-31 |
| Yan, Shannon; Wen, Jin-Der; Bustamante, Carlos et al. (2015) Ribosome excursions during mRNA translocation mediate broad branching of frameshift pathways. Cell 160:870-881 |
| Shukla, C; Koch, L G; Britton, S L et al. (2015) Contribution of regional brain melanocortin receptor subtypes to elevated activity energy expenditure in lean, active rats. Neuroscience 310:252-67 |
| Tinoco Jr, Ignacio (2014) Fun and games in Berkeley: the early years (1956-2013). Annu Rev Biophys 43:1-17 |
| Kaiser, Christian M; Tinoco Jr, Ignacio (2014) Probing the mechanisms of translation with force. Chem Rev 114:3266-80 |
| Kim, Hee-Kyung; Liu, Fei; Fei, Jingyi et al. (2014) A frameshifting stimulatory stem loop destabilizes the hybrid state and impedes ribosomal translocation. Proc Natl Acad Sci U S A 111:5538-43 |
| Liu, Tingting; Kaplan, Ariel; Alexander, Lisa et al. (2014) Direct measurement of the mechanical work during translocation by the ribosome. Elife 3:e03406 |
| Tinoco Jr, Ignacio; Kim, Hee-Kyung; Yan, Shannon (2013) Frameshifting dynamics. Biopolymers 99:1147-66 |
| Qu, Xiaohui; Lancaster, Laura; Noller, Harry F et al. (2012) Ribosomal protein S1 unwinds double-stranded RNA in multiple steps. Proc Natl Acad Sci U S A 109:14458-63 |
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