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Use of a Recombinant Mosquito Densovirus As a Gene Delivery Vector for the Functional Analysis of Genes in Mosquito Larvae
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Aedes aegypti encodes an ATPase-active RUVBL1/2 complex.
Natália G Quel1, Leonardo T Rosa2, Larissa M Antonio1
1Institute of Chemistry, University of Campinas (UNICAMP), Campinas, SP 13083-970, Brazil; National Institute of Science and Technology for Bioimage and Structural Biology INBEB, Brazil.
International Journal of Biological Macromolecules
|June 12, 2025
Summary
The RUVBL1/2 complex from Aedes aegypti forms a dodecamer with ATPase activity, but requires both proteins to function, unlike the human version. This study provides structural insights into this essential protein complex in a disease vector.
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- RUVBL1 and RUVBL2 proteins are crucial for DNA repair and chromatin homeostasis in eukaryotes, functioning as ATPases and chaperones.
- Previous studies on human and yeast RUVBL1/2 orthologs show species-specific differences in dynamics, interactions, and ATPase activity.
- Investigating diverse eukaryotic orthologs is key to understanding conserved RUVBL1/2 structure and function.
Purpose of the Study:
- To identify and characterize RUVBL1 and RUVBL2 proteins from Aedes aegypti, a significant vector for viral diseases.
- To determine the structural and functional properties of the Aedes aegypti RUVBL1/2 (AaRUVBL1/2) complex.
- To compare the functional and structural characteristics of AaRUVBL1/2 with its human and yeast counterparts.
Main Methods:
- Bioinformatic analysis of the Aedes aegypti genome to identify RUVBL1/2 orthologs.
- Purification and characterization of recombinant AaRUVBL1/2 complex, including ATPase activity assays and mutational analysis.
- Structural determination using Small-Angle X-ray Scattering (SAXS) and Cryo-Electron Microscopy (Cryo-EM).
Main Results:
- Identified and characterized AaRUVBL1 and AaRUVBL2, which assemble into a hetero-dodecameric complex with ATPase activity.
- Demonstrated that AaRUVBL1/2 ATPase activity is dependent on the presence of both subunits, contrasting with human RUVBL1/2.
- Structural analysis revealed a barrel-shaped dodecamer (~16 nm) with significant conformational heterogeneity within and between hexameric rings.
Conclusions:
- The Aedes aegypti RUVBL1/2 complex is a bona fide AAA+ ATPase with a unique requirement for both subunits for activity.
- Structural insights reveal a heterogeneous dodecameric complex, providing a foundation for understanding its function in this important insect vector.
- This study represents the second structurally and functionally characterized RUVBL complex within the Animalia phylum, expanding the comparative understanding of these vital proteins.
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