Full molecular trajectories of RNA polymerase at single base-pair resolution

Journal Article · 19 January 2018 · Proceedings of the National Academy of Sciences of the United States of America

DOI: https://doi.org/10.1073/pnas.1719906115 · OSTI ID:1417527

Righini, Maurizio ^[1]; Lee, Antony ^[2]; Cañari-Chumpitaz, Cristhian ^[1]; Lionberger, Troy ^[3]; Gabizon, Ronen ^[4]; Coello, Yves  ^[5]; Tinoco, Jr., Ignacio ^[6]; Bustamante, Carlos  ^[7]

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1. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, Department of Chemistry, University of California, Berkeley, CA 94720,
2. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, Department of Physics, University of California, Berkeley, CA 94720,
3. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, CA 94720,, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720,
4. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, CA 94720,
5. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, CA 94720,, Departamento de Ciencias, Sección Química, Pontificia Universidad Católica del Perú, Lima 32, Peru,
6. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,
7. Jason L. Choy Laboratory of Single-Molecule Biophysics, University of California, Berkeley, CA 94720,, Department of Chemistry, University of California, Berkeley, CA 94720,, California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, CA 94720,, Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720,, Kavli Energy Nanoscience Institute, University of California, Berkeley, CA 94720

Significance Optical tweezers enable scientists to follow the dynamics of molecular motors at high resolution. The ability to discern a motor’s discrete steps reveals important insights on its operation. Some motors operate at the scale of angstroms, rendering the observation of their steps extremely challenging. In some cases, such small steps have been observed sporadically; however, the full molecular trajectories of steps and intervals between steps remain elusive due to instrumental noise. Here, we eliminate the main source of noise of most high-resolution dual-trap optical tweezers and developed both a single-molecule assay and a self-learning algorithm to uncover the full trajectories of such a motor: RNA polymerase. Using this method, a whole new set of experiments becomes possible.

• Sponsoring Organization: USDOE
• Grant/Contract Number: AC02-05CH11231
• OSTI ID: 1417527
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 6 Vol. 115; ISSN 0027-8424
• Publisher: Proceedings of the National Academy of SciencesCopyright Statement
• Country of Publication: United States
• Language: English