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Related Concept Videos

DNA Topoisomerases02:02

DNA Topoisomerases

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...
Atomic Force Microscopy01:08

Atomic Force Microscopy

Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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Studying the Cytoskeleton01:17

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...

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High-Speed Atomic Force Microscopy Imaging of DNA Three-Point-Star Motif Self Assembly Using Photothermal Off-Resonance Tapping
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Human topoisomerase II-DNA interaction study by using atomic force microscopy.

Livan Alonso-Sarduy1, Charles Roduit, Giovanni Dietler

  • 1Laboratoire de Physique de la Matière Vivante, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland. livan.alonso@epfl.ch

FEBS Letters
|September 13, 2011
PubMed
Summary
This summary is machine-generated.

Human topoisomerases II (Topo II) prefer binding to DNA crossovers, influencing DNA topology. Atomic force microscopy revealed preferred binding angles and visualized DNA unknotting dynamics.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • Type II topoisomerases (Topo II) are essential enzymes that regulate DNA topology through ATP-dependent double-strand breaks and passages.
  • Understanding the precise mechanisms of Topo II binding and action is crucial for comprehending DNA replication, transcription, and repair.

Purpose of the Study:

  • To investigate the preferred binding sites and geometric preferences of human Topo II on DNA using atomic force microscopy (AFM).
  • To visualize the dynamic process of DNA unknotting mediated by human Topo II.

Main Methods:

  • Atomic force microscopy (AFM) was employed to directly visualize the interaction of human Topo II with DNA molecules.
  • Supercoiled and knotted DNA substrates were used to study Topo II's relaxation and unknotting activities.

Main Results:

  • Human Topo II demonstrated a preference for binding to DNA crossovers, suggesting a specific recognition mechanism.
  • A favored binding geometry with angles between 80-90° at DNA crossings was observed, indicating chiral discrimination.
  • AFM imaging successfully visualized the dynamic process of Topo II unknotting complex DNA structures.

Conclusions:

  • Human Topo II exhibits specific binding preferences and geometric requirements at DNA crossovers.
  • AFM provides valuable insights into the dynamic action of Topo II in altering DNA topology.
  • These findings contribute to a deeper understanding of Topo II function in managing DNA structure and dynamics.