Title
Full molecular trajectories of RNA polymerase at single base-pair resolution
Date Issued
06 February 2018
Access level
open access
Resource Type
journal article
Author(s)
Righini M.
Lee A.
Cañari-Chumpitaz C.
Lionberger T.
Gabizon R.
Tinoco I.
Publisher(s)
National Academy of Sciences
Abstract
In recent years, highly stable optical tweezers systems have enabled the characterization of the dynamics of molecular motors at very high resolution. However, the motion of many motors with angstrom-scale dynamics cannot be consistently resolved due to poor signal-to-noise ratio. Using an acousto-optic deflector to generate a "time-shared" dual-optical trap, we decreased low-frequency noise by more than one order of magnitude compared with conventional dual-trap optical tweezers. Using this instrument, we implemented a protocol that synthesizes single base-pair trajectories, which are used to test a Large State Space Hidden Markov Model algorithm to recover their individual steps. We then used this algorithm on real transcription data obtained in the same instrument to fully uncover the molecular trajectories of Escherichia coli RNA polymerase. We applied this procedure to reveal the effect of pyrophosphate on the distribution of dwell times between consecutive polymerase steps.
Start page
1286
End page
1291
Volume
115
Issue
6
Language
English
OCDE Knowledge area
Física atómica, molecular y química
Subjects
Scopus EID
2-s2.0-85041498219
PubMed ID
Source
Proceedings of the National Academy of Sciences of the United States of America
ISSN of the container
0027-8424
Sponsor(s)
ACKNOWLEDGMENTS. We dedicate this manuscript to the memory of Ignacio Tinoco Jr. We thank Matthew J. Comstock and Charles Wickersham for advice and help in setting up the time-shared trap and the members of the C.B. laboratory for helpful discussions. This work was supported by Howard Hughes Medical Institute (C.B.), NIH Grants R01GM032543 (to C.B.) and R01GM071552 (to C.B.), and the US Department of Energy Office of Basic Energy Sciences Nanomachine Program under Contract DE-AC02-05CH11231 (to C.B.).
Sources of information:
Directorio de Producción Científica
Scopus