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

Maurizio Righini, Antony Lee, Cristhian Cañari-Chumpitaz, Troy A. Lionberger, Ronen Gabizon, Yves Coello, Ignacio Tinoco, Carlos Bustamante

Producción científica: Contribución a una revistaArtículorevisión exhaustiva

23 Citas (Scopus)

Resumen

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.
Idioma originalEspañol
Páginas (desde-hasta)1286-1291
Número de páginas6
PublicaciónProceedings of the National Academy of Sciences of the United States of America
Volumen115
EstadoPublicada - 6 feb. 2018

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