Single-molecule real-time 3D imaging of the transcription cycle by modulation interferometry Journal Article

   


Authors: Wang, G.; Hauver, J.; Thomas, Z.; Darst, S. A.; Pertsinidis, A.
Article Title: Single-molecule real-time 3D imaging of the transcription cycle by modulation interferometry
Abstract: Many essential cellular processes, such as gene control, employ elaborate mechanisms involving the coordination of large, multi-component molecular assemblies. Few structural biology tools presently have the combined spatial-temporal resolution and molecular specificity required to capture the movement, conformational changes, and subunit association-dissociation kinetics, three fundamental elements of how such intricate molecular machines work. Here, we report a 3D single-molecule super-resolution imaging study using modulation interferometry and phase-sensitive detection that achieves <2 nm axial localization precision, well below the few-nanometer-sized individual protein components. To illustrate the capability of this technique in probing the dynamics of complex macromolecular machines, we visualize the movement of individual multi-subunit E. coli RNA polymerases through the complete transcription cycle, dissect the kinetics of the initiation-elongation transition, and determine the fate of σ70 initiation factors during promoter escape. Modulation interferometry sets the stage for single-molecule studies of several hitherto difficult-to-investigate multi-molecular transactions that underlie genome regulation. © 2016 Elsevier Inc.
Keywords: transcription; rna polymerase; sigma factor; single-molecule; holoenzyme; nuclear pore complex; interferometry; transcription cycle; axial localization; super-resolution imaging
Journal Title: Cell
Volume: 167
Issue: 7
ISSN: 0092-8674
Publisher: Cell Press  
Date Published: 2016-12-15
Start Page: 1839
End Page: 1852.e21
Language: English
DOI: 10.1016/j.cell.2016.11.032
PROVIDER: scopus
PUBMED: 27984731
PMCID: PMC5444671
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85006262143&doi=10.1016%2fj.cell.2016.11.032&partnerID=40&md5=dea4fced0359f0fe8eb6e192af1702d1
Notes: Article -- Export Date: 3 January 2017 -- Source: Scopus
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MSK Authors
  1. Alexandros Pertsinidis
    16 Pertsinidis
  2. Guanshi Wang
    6 Wang
  3. Jesse J. Hauver
    3 Hauver
  4. Zachary Thomas
    2 Thomas
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