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Updated: May 29, 2026

Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies
Published on: November 28, 2017
Structural and functional insights into a dodecameric molecular machine - the RuvBL1/RuvBL2 complex
Sabine Gorynia1, Tiago M Bandeiras, Filipa G Pinho
1Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
The RuvBL1 and RuvBL2 complex structure reveals a dodecameric ring, essential for DNA repair and cancer regulation. Domain II truncation enhances ATP consumption and DNA unwinding activity, suggesting cofactor-mediated regulation.
Area of Science:
- Biochemistry
- Molecular Biology
- Structural Biology
Background:
- RuvBL1 and RuvBL2 are conserved AAA(+) ATPases crucial for DNA repair, chromatin remodeling, and transcriptional regulation.
- Overexpression of RuvBL1 and RuvBL2 is linked to various cancers, where they interact with oncogenic factors like β-catenin and c-Myc.
Purpose of the Study:
- To determine the three-dimensional crystal structure of the human RuvBL complex with a truncated domain II.
- To investigate the impact of domain II truncation on the helicase activity and ATP consumption of RuvBL1 and RuvBL2.
Main Methods:
- X-ray crystallography to solve the 3D structure of the RuvBL complex.
- Small-angle X-ray scattering (SAXS) to confirm the quaternary structure.
- Biochemical assays to measure ATP consumption and DNA unwinding activity.
Main Results:
- The crystal structure revealed a dodecameric complex of two heterohexameric rings with alternating RuvBL1 and RuvBL2 monomers.
- Truncation of domain II significantly increased ATP consumption and enhanced DNA unwinding activity.
- Domain II was found to auto-inhibit the DNA unwinding activity of the human RuvBL proteins.
Conclusions:
- The dodecameric structure of the RuvBL complex is essential for its function.
- Domain II plays a regulatory role in RuvBL activity, acting as an auto-inhibitory domain.
- Cofactor-mediated conformational changes involving domain II likely regulate the in vivo activity of RuvBL proteins as therapeutic targets.
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