Evidence map›Paper›PMID 39484392›Full record

ArticlebioRxiv : the preprint server for biology2024

A high throughput single molecule platform to study DNA supercoiling effect on protein-DNA interactions.

Huijin Lee, Jihee Hwang, Fahad Rashid, James A London, Richard Fishel, James M Berger, Sua Myong, Taekjip Ha

Abstract readPreprint
In one paragraph

Article in bioRxiv : the preprint server for biology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

0numbers the graph read from it
0cells of the map it votes in
0citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

5 · Who and what money

Authors and funding

8 authors.

Huijin LeeDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland, 21205, USA.ORCID 0009-0007-1261-0139
Jihee HwangPrograms in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, Massachusetts, 02115, USA.
Fahad RashidDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland, 21205, USA.
James A LondonDepartment of Cancer Biology and Genetics, The Ohio State University Wexner Medical Center, Columbus, Ohio, 43210, USA.
Richard FishelDepartment of Cancer Biology and Genetics, The Ohio State University Wexner Medical Center, Columbus, Ohio, 43210, USA.
James M BergerDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland, 21205, USA.ORCID 0000-0003-0666-1240
Sua MyongDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland, 21205, USA.
Taekjip HaDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins School of Medicine, Baltimore, Maryland, 21205, USA.ORCID 0000-0003-2195-6258

Funding

The Human Mismatch Repair Proteins and CarcinogenesisR01CA067007 · NCI · UNIVERSITY OF VERMONT &ST AGRIC COLLEGE · PI Richard Fishel · 1995 to 2026
$9.6M
Understanding and exploiting DNA topoisomerases in cancer biologyR35CA263778 · NCI · JOHNS HOPKINS UNIVERSITY · PI James M. Berger · 2021 to 2026
$5.0M
Mechanistic Studies of Replication Initiation in ProkaryotesR37GM071747 · NIGMS · JOHNS HOPKINS UNIVERSITY · PI James M. Berger · 2017 to 2026
$4.4M
Single Molecule Studies of Nucleic Acids RemodelingR35GM122569 · NIGMS · JOHNS HOPKINS UNIVERSITY · PI Taekjip Ha · 2017 to 2026
$3.5M
Gene Expression Modulated by G4, R-loop and DNA Supercoiling (Supplement for Equipment)R01GM149729 · NIGMS · BOSTON CHILDREN'S HOSPITAL · PI Sua Myong · 2023 to 2026
$1.6M
NCI NIH HHS R01 CA067007NCI NIH HHS R35 CA263778NIGMS NIH HHS R01 GM149729NIGMS NIH HHS R35 GM122569NIGMS NIH HHS R37 GM071747
6 · The paper itself

Abstract

DNA supercoiling significantly influences DNA metabolic pathways. To examine its impact on DNA-protein interactions at the single-molecule level, we developed a highly efficient and reliable protocol to modify plasmid DNA at specific sites, allowing us to label plasmids with fluorophores and biotin. We then induced negative and positive supercoiling in these plasmids using gyrase and reverse gyrase, respectively. Comparing supercoiled DNA with relaxed circular DNA, we assessed the effects of supercoiling on CRISPR-Cas9 and mismatch repair protein MutS. We found that negative DNA supercoiling exacerbates off-target effects in DNA unwinding by Cas9. For MutS, we observed both negative and positive DNA supercoiling enhances the binding interaction between MutS and a mismatched base pair but does not affect the rate of ATP-induced sliding clamp formation. These findings not only underscore the versatility of our protocol but also opens new avenues for exploring the intricate dynamics of protein-DNA interactions under the influences of supercoiling.

Identifiers

PMID39484392
PMCPMC11527113

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.