DNA sequence encodes the position of DNA supercoils
Abstract
The three-dimensional organization of DNA is increasingly understood to play a decisive role in vital cellular processes. Many studies focus on the role of DNA-packaging proteins, crowding, and confinement in arranging chromatin, but structural information might also be directly encoded in bare DNA itself. Here we visualize plectonemes (extended intertwined DNA structures formed upon supercoiling) on individual DNA molecules. Remarkably, our experiments show that the DNA sequence directly encodes the structure of supercoiled DNA by pinning plectonemes at specific sequences. We develop a physical model that predicts that sequence-dependent intrinsic curvature is the key determinant of pinning strength and demonstrate this simple model provides very good agreement with the data. Analysis of several prokaryotic genomes indicates that plectonemes localize directly upstream of promoters, which we experimentaly confirm for selected promotor sequences. Our findings reveal a hidden code in the genome that helps to spatially organize the chromosomal DNA.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. The previously published genome data for E. coli used in Figure 4B can be accessed here http://regulondb.ccg.unam.mx/menu/download/datasets/files/PromoterSet.txt; V. cholerae here http://www.pnas.org/highwire/filestream/618514/field_highwire_adjunct_files/2/pnas.1500203112.sd02.xlsx; B. methanolicus here https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4342826/bin/12864_2015_1239_MOESM2_ESM.xlsx; M. tuberculosis here https://ars.els-cdn.com/content/image/1-s2.0-S2211124713006153-mmc2.xlsx; and C. crescentus here https://doi.org/10.1371/journal.pgen.1004831.s012. The previously published genome data for D. melanogaster, C. elegans, A. thaliana, S. cerevisiae, and S. pombe used in Figure 4E can be accessed using the Eukaryotic Promotor Database (https://epd.vital-it.ch).
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Whole genome sequence data: E. ColiNCBI Genome, NC_000913.3.
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Whole genome sequence data: S. cerevisiase (Chr I)NCBI Nucleotide, NC_001133.9.
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Whole genome sequence data: S. cerevisiase (Chr II)NCBI Nucleotide, NC_001134.8.
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Whole genome sequence data: S. cerevisiase (Chr III)NCBI Nucleotide, NC_001135.5.
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Whole genome sequence data: S. cerevisiase (Chr IV)NCBI Nucleotide, NC_001136.10.
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Whole genome sequence data: S. cerevisiase (Chr V)NCBI Nucleotide, NC_001137.3.
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Whole genome sequence data: S. cerevisiase (Chr VI)NCBI Nucleotide, NC_001138.5.
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Whole genome sequence data: S. cerevisiase (Chr VII)NCBI Nucleotide, NC_001139.9.
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Whole genome sequence data: S. cerevisiase (Chr VIII)NCBI Nucleotide, NC_001140.6.
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Whole genome sequence data: S. cerevisiase (Chr IX)NCBI Nucleotide, NC_001141.2.
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Whole genome sequence data: S. cerevisiase (Chr X)NCBI Nucleotide, NC_001142.9.
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Whole genome sequence data: S. cerevisiase (Chr XI)NCBI Nucleotide, NC_001143.9.
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Whole genome sequence data: S. cerevisiase (Chr XII)NCBI Nucleotide, NC_001144.5.
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Whole genome sequence data: S. cerevisiase (Chr XIII)NCBI Nucleotide, NC_001145.3.
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Whole genome sequence data: S. cerevisiase (Chr XIV)NCBI Nucleotide, NC_001146.8.
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Whole genome sequence data: S. cerevisiase (Chr XV)NCBI Nucleotide, NC_001147.6.
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Whole genome sequence data: S. cerevisiase (Chr XVI)NCBI Nucleotide, NC_001148.4.
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Whole genome sequence data: S. cerevisiaseNCBI Nucleotide, NC_001224.1.
Article and author information
Author details
Funding
H2020 European Research Council (669598)
- Cees Dekker
The Netherlands Organization for Scientific Research (the Frontiers of Nanoscience program)
- Elio Abbondanzieri
H2020 European Research Council (304284)
- Elio Abbondanzieri
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Michael T Laub, Massachusetts Institute of Technology, United States
Version history
- Received: March 10, 2018
- Accepted: December 6, 2018
- Accepted Manuscript published: December 7, 2018 (version 1)
- Version of Record published: December 20, 2018 (version 2)
Copyright
© 2018, Kim et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Further reading
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RNA polymerase II (RNAPII) transcription initiates bidirectionally at many human protein-coding genes. Sense transcription usually dominates and leads to messenger RNA production, whereas antisense transcription rapidly terminates. The basis for this directionality is not fully understood. Here, we show that sense transcriptional initiation is more efficient than in the antisense direction, which establishes initial promoter directionality. After transcription begins, the opposing functions of the endonucleolytic subunit of Integrator, INTS11, and cyclin-dependent kinase 9 (CDK9) maintain directionality. Specifically, INTS11 terminates antisense transcription, whereas sense transcription is protected from INTS11-dependent attenuation by CDK9 activity. Strikingly, INTS11 attenuates transcription in both directions upon CDK9 inhibition, and the engineered recruitment of CDK9 desensitises transcription to INTS11. Therefore, the preferential initiation of sense transcription and the opposing activities of CDK9 and INTS11 explain mammalian promoter directionality.
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