Properties of Oligomeric Interaction of the Cytomegalovirus Core Nuclear Egress Complex (NEC) and Its Sensitivity to an NEC Inhibitory Small Molecule
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Plasmid Constructs
2.3. Antiviral Compounds
2.4. Antibodies
2.5. PEI Transfection and Coimmunoprecipitation (CoIP)
2.6. In Vitro NEC Assembly Assay
2.7. Virus Infection
2.8. FuGENE® Transient Transfection, Indirect Immunofluorescence Assay and Confocal Laser-Scanning Microscopy
2.9. Bioinformatic Methods
3. Results
3.1. Functional Principle of the HCMV-Specific Nuclear Egress Complex and Generation of Test Constructs to Analyze Its Oligomeric Binding Properties
3.2. An In Vitro NEC Assembly Assay Strongly Supports the Idea of Oligomeric HCMV pUL50-pUL53 Interaction
3.3. Confirmation of Conditions of Oligomeric pUL50-pUL53 Interaction Using the Standard CoIP Approach
3.4. Interface Analysis of the Hexameric NEC and Computational Investigation of pUL50 Conformational Stability
3.5. Blocking of In Vitro Assembly-Based Oligomeric pUL50-pUL53 Interaction by an NEC-Directed Inhibitor
3.6. Interference of the Inhibitory Small Molecule MBM with NEC Nuclear Rim Formation and Viral Nuclear Egress Activity
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Kicuntod, J.; Alkhashrom, S.; Häge, S.; Diewald, B.; Müller, R.; Hahn, F.; Lischka, P.; Sticht, H.; Eichler, J.; Marschall, M. Properties of Oligomeric Interaction of the Cytomegalovirus Core Nuclear Egress Complex (NEC) and Its Sensitivity to an NEC Inhibitory Small Molecule. Viruses 2021, 13, 462. https://doi.org/10.3390/v13030462
Kicuntod J, Alkhashrom S, Häge S, Diewald B, Müller R, Hahn F, Lischka P, Sticht H, Eichler J, Marschall M. Properties of Oligomeric Interaction of the Cytomegalovirus Core Nuclear Egress Complex (NEC) and Its Sensitivity to an NEC Inhibitory Small Molecule. Viruses. 2021; 13(3):462. https://doi.org/10.3390/v13030462
Chicago/Turabian StyleKicuntod, Jintawee, Sewar Alkhashrom, Sigrun Häge, Benedikt Diewald, Regina Müller, Friedrich Hahn, Peter Lischka, Heinrich Sticht, Jutta Eichler, and Manfred Marschall. 2021. "Properties of Oligomeric Interaction of the Cytomegalovirus Core Nuclear Egress Complex (NEC) and Its Sensitivity to an NEC Inhibitory Small Molecule" Viruses 13, no. 3: 462. https://doi.org/10.3390/v13030462