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Updated: Aug 4, 2025

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C

Deciphering High-Resolution 3D Chromatin Organization via Capture Hi-C

Published on: October 14, 2022

DNA segment capture by Smc5/6 holocomplexes.

Michael Taschner¹, Stephan Gruber²

¹Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland.

Nature Structural & Molecular Biology

|April 3, 2023

Summary

This summary is machine-generated.

Yeast Smc5/6 complexes load ATP-dependently onto DNA, requiring the Nse5/6 subcomplex to open the kleisin neck gate. This mechanism entraps DNA, potentially explaining how structural maintenance of chromosomes (SMC) complexes extrude DNA loops.

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Selective Capture of 5-hydroxymethylcytosine from Genomic DNA

Selective Capture of 5-hydroxymethylcytosine from Genomic DNA

Published on: October 5, 2012

Area of Science:

Molecular Biology
Chromatin Biology
Biochemistry

Background:

Structural Maintenance of Chromosomes (SMC) complexes are crucial for chromosome folding and segregation in eukaryotes, primarily through DNA loop extrusion.
The precise mechanism by which SMC complexes interact with DNA to extrude loops remains incompletely understood.
The Smc5/6 complex plays specialized roles in DNA repair and the resolution of aberrant DNA junctions.

Purpose of the Study:

To reconstitute and characterize the ATP-dependent DNA loading mechanism of yeast Smc5/6 complexes.
To elucidate the role of the Nse5/6 subcomplex in Smc5/6 DNA engagement and loop formation.
To understand how DNA is topologically managed within the Smc5/6 complex during loading and potential extrusion.

Main Methods:

Reconstitution of functional yeast Smc5/6 complexes in vitro.
Biochemical assays to monitor ATP-dependent DNA binding and loading.
Topological analysis of plasmid DNA interacting with the Smc5/6 complex and its subcomplexes.

Main Results:

ATP-dependent DNA loading by yeast Smc5/6 rings was successfully reconstituted.
The Nse5/6 subcomplex is essential for DNA loading, acting by opening the kleisin neck gate.
Plasmid DNA is topologically entrapped within the kleisin and two SMC subcompartments, but not the full SMC compartment, indicating a specific DNA-holding mechanism.

Conclusions:

The Nse5/6 subcomplex acts as a gatekeeper for DNA entry into the Smc5/6 complex.
The observed topological entrapment explains how Smc5/6 holds a looped DNA segment, with the kleisin locking it during closure.
This mechanism of DNA segment capture may provide the power stroke for subsequent DNA extrusion, offering a unifying principle for SMC complex function.