Supercoiling

Supercoiling is the three-dimensional twisting of a DNA molecule around its own axis or into higher-order coils, a structural feature that helps organize and regulate genetic material. It arises when the DNA double helix becomes overwound or underwound, changing its linking number and creating torsional strain; enzymes called topoisomerases relieve or introduce this strain by transiently breaking and resealing DNA strands. In biology, supercoiling influences chromosome compaction, replication, transcription, and recombination by altering DNA accessibility. Measuring or manipulating these topological states helps researchers study genome organization, enzyme activity, and mechanisms of gene regulation.

Supercoiling - Related Videos

Education

JoVE Core - Molecular Biology

DNA Topoisomerases

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2020

Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA. Types and Mechanism of action Topoisomerases are divided into two main types. Type I...

Research

JoVE Journal - Genetics

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

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2016

Targeted DNA damage can be achieved by tethering a DNA damaging agent to a triplex-forming oligonucleotide (TFO). Using modified TFOs, DNA damage-specific protein association, and DNA topology modification can be studied in human cells by the utilization of modified chromatin immunoprecipitation assays and DNA supercoiling assays described herein.

Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase

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Cited by 10 •

2017

Recombinant prototype foamy virus integrase protein is often contaminated with a bacterial nuclease during purification. This method identifies nuclease contamination and removes it from the final preparation of the enzyme.

Alkaline Single Cell Gel Electrophoresis: A Sensitive Technique for Quantitative Estimation of DNA Damage in Individual Cancer Cells Following Chemotherapy

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2025

In this video, we demonstrate an alkaline single-cell electrophoresis assay to quantify both single- and double-strand DNA breaks in doxorubicin-treated cells. Alkaline conditions disrupt the hydrogen bonds between DNA base pairs, resulting in the complete unwinding and separation of DNA strands. This allows differential migration of undamaged and damaged DNA fragments in agarose gel upon electrophoresis, revealing a comet-shaped structure of slowly migrating undamaged DNA forming the circular...

Bacterial Transformation: The Heat Shock Method

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2023

Transformation is the process that occurs when a cell ingests foreign DNA from its surroundings. Transformation can occur in nature in certain types of bacteria. In molecular biology, transformation is artificially reproduced in the lab via the creation of pores in bacterial cell membranes. Bacterial cells that are able to take up DNA from the environment are called competent cells. In the laboratory, bacterial cells can be made competent and DNA subsequently introduced by a procedure called...

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