Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway
Abstract
:1. Introduction
2. Results and Discussion
2.1. TEM and Cryo-EM Analysis Showed Differential Shapes of Enveloped WSSV Virions and Nucleocapsids
2.2. Three-Dimensional Volume Reconstruction of WSSV Nucleocapsid by Cryo-Electron Tomography (Cryo-ET)
2.3. Proposed WSSV Genomic DNA Translocation through the Cap End of WSSV Nucleocapsids
2.4. Cryo-EM Map of the Middle Part of the WSSV Nucleocapsid (M-NC) Revealed a Unique Architecture with C14 Symmetry
2.5. WSSV VP664 Is the Main Component of the 14 NUs of WSSV Nucleocapsid
2.6. Proposed Model for WSSV Morphogenesis
3. Materials and Methods
3.1. Preparation of WSSV Inoculum
3.2. Purification of WSSV Virions and Nucleocapsids
3.3. Cryo-EM Sample Vitrification
3.4. Data Acquisition
3.5. Image Processing
3.6. Ultrastructural Observations by Transmission Electron Microscopy
3.7. Localization of WSSV Major Nucleocapsid Protein VP664 on the Nucleocapsid Rings by Immunoelectron Microscopy (IEM)
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Type of WSSV Nucleocapsid | Narrow Type | Wide Type |
---|---|---|
Electron microscope | Talos Arctica | Talos Arctica |
Detector | Falcon Ⅲ | Falcon Ⅲ |
Magnification | 73,000× | 73,000× |
Voltage(kV) | 200 | 200 |
Electron exposure (e−/Å2) | 48 | 48 |
Exposure (s) | 2.5 | 2.5 |
Frames (no.) | 50 | 50 |
Defocus range (um) | −0.5~−2.5 | −0.5~−2.5 |
Pixel size (Å) | 2.76 | 2.76 |
Symmetry imposed | C14 | C14 |
Initial particles | 55,723 | 55,723 |
Final particles | 15,279 | 15,243 |
Map resolution (Å) | 5.52 | 5.52 |
FSC threshold | 0.143 | 0.143 |
Software | Relion and cisTEM | Relion and cisTEM |
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Huang, H.-J.; Tang, S.-L.; Chang, Y.-C.; Wang, H.-C.; Ng, T.H.; Garmann, R.F.; Chen, Y.-W.; Huang, J.-Y.; Kumar, R.; Chang, S.-H.; et al. Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway. Int. J. Mol. Sci. 2023, 24, 7525. https://doi.org/10.3390/ijms24087525
Huang H-J, Tang S-L, Chang Y-C, Wang H-C, Ng TH, Garmann RF, Chen Y-W, Huang J-Y, Kumar R, Chang S-H, et al. Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway. International Journal of Molecular Sciences. 2023; 24(8):7525. https://doi.org/10.3390/ijms24087525
Chicago/Turabian StyleHuang, Hui-Ju, Sen-Lin Tang, Yuan-Chih Chang, Hao-Ching Wang, Tze Hann Ng, Rees F. Garmann, Yu-Wen Chen, Jiun-Yan Huang, Ramya Kumar, Sheng-Hsiung Chang, and et al. 2023. "Multiple Nucleocapsid Structural Forms of Shrimp White Spot Syndrome Virus Suggests a Novel Viral Morphogenetic Pathway" International Journal of Molecular Sciences 24, no. 8: 7525. https://doi.org/10.3390/ijms24087525