Single-molecule nanoscopy elucidates RNA polymerase II transcription at single genes in live cells Journal Article

   


Authors: Li, J.; Dong, A.; Saydaminova, K.; Chang, H.; Wang, G.; Ochiai, H.; Yamamoto, T.; Pertsinidis, A.
Article Title: Single-molecule nanoscopy elucidates RNA polymerase II transcription at single genes in live cells
Abstract: Transforming the vast knowledge from genetics, biochemistry, and structural biology into detailed molecular descriptions of biological processes inside cells remains a major challenge—one in sore need of better imaging technologies. For example, transcription involves the complex interplay between RNA polymerase II (Pol II), regulatory factors (RFs), and chromatin, but visualizing these dynamic molecular transactions in their native intracellular milieu remains elusive. Here, we zoom into single tagged genes using nanoscopy techniques, including an active target-locking, ultra-sensitive system that enables single-molecule detection in addressable sub-diffraction volumes, within crowded intracellular environments. We image, track, and quantify Pol II with single-molecule resolution, unveiling its dynamics during the transcription cycle. Further probing multiple functionally linked events—RF-chromatin interactions, Pol II dynamics, and nascent transcription kinetics—reveals detailed operational parameters of gene-regulatory mechanisms hitherto-unseen in vivo. Our approach sets the stage for single-molecule studies of complex molecular processes in live cells. Key limitations of imaging single copies of highly abundant factors at specific intracellular sites of activity are overcome by an active target-locking system that achieves long-term tracking at single-molecule resolution. © 2019 Elsevier Inc.
Journal Title: Cell
Volume: 178
Issue: 2
ISSN: 0092-8674
Publisher: Cell Press  
Date Published: 2019-07-11
Start Page: 491
End Page: 506.e28
Language: English
DOI: 10.1016/j.cell.2019.05.029
PUBMED: 31155237
PROVIDER: scopus
PMCID: PMC6675578
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85068374322&doi=10.1016%2fj.cell.2019.05.029&partnerID=40&md5=9a5e2376abfd8ac584fbaf983c448db6
Notes: Source: Scopus
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MSK Authors
  1. Alexandros Pertsinidis
    16 Pertsinidis
  2. Ankun Dong
    2 Dong
  3. Guanshi Wang
    6 Wang
  4. Kamola Saydaminova
    1 Saydaminova
  5. Hill Zheng-Hao Chang
    1 Chang
  6. Jieru Li
    6 Li
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