期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 113, 期 45, 页码 12733-12738出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1602764113
关键词
Pol II; transcription; single-molecule; optical tweezers; internucleosomal distance
资金
- CIPSM
- NIM
- QBM
- Advanced Investigator Grant TRANSIT of the European Research Council
- Volkswagen Foundation
- European Research Council [281903]
- Innovative Training Networks (ITN) from the European Union [281903, 641639]
- Human Frontier Science Program [RGP0023/2014]
- Deutsche Forschungsgemeinschaft (DFG) [SFB646, SFB960, SFB1064, GRK1721]
- Max Planck Society
- European Research Council (ERC) [281903] Funding Source: European Research Council (ERC)
In eukaryotes, gene expression depends on chromatin organization. However, how chromatin affects the transcription dynamics of individual RNA polymerases has remained elusive. Here, we use dual trap optical tweezers to study single yeast RNA polymerase II (Pol II) molecules transcribing along a DNA template with two nucleosomes. The slowdown and the changes in pausing behavior within the nucleosomal region allow us to determine a drift coefficient,., which characterizes the ability of the enzyme to recover from a nucleosomal backtrack. Notably, chi can be used to predict the probability to pass the first nucleosome. Importantly, the presence of a second nucleosome changes chi in a manner that depends on the spacing between the two nucleosomes, as well as on their rotational arrangement on the helical DNA molecule. Our results indicate that the ability of Pol II to pass the first nucleosome is increased when the next nucleosome is turned away from the first one to face the opposite side of the DNA template. These findings help to rationalize how chromatin arrangement affects Pol II transcription dynamics.
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