Abstract
The membrane-proximal external region (MPER) of Lassa virus (LASV) glycoprotein complex (GPC) is critical in modulating its functionality. Till now, the high-resolution structure of the intact GPC, including MPER is not available. In this study, we used alanine substitution to scan all 16 residues located in LASV MPER. Western blotting and quantification fusion assay showed that the residues located at the C terminus of the HR2 (M414 and L415) and N terminus of the MPER (K417 and Y419) are critical for GPC-mediated membrane fusion function. Furthermore, cell surface biotinylation experiments revealed that M414A, K417A and Y419A expressed similar levels as WT, whereas L415A mutant led to a reduction of mature GPC on the cell surface. Moreover, substitution of these residues with the similar residue such as M414L, L415I, K417R and Y419F would partly compensate the loss of the fusion activity caused by the alanine mutant in these sites. Results from this study showed that several key residues in the MPER region are indispensable to promote the conformational changes that drive fusion events and shed light on the structure analysis of LASV GPC and anti-LASV therapeutics.
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Acknowledgements
We thank the Center for Instrumental Analysis and Metrology, Core Facility and Technical Support, and Center for Animal Experiment, Wuhan Institute of Virology, for providing technical assistance. This work was supported by the National Key Research and Development Program of China (2018YFA0507204), the National Natural Sciences Foundation of China (31670165), Wuhan National Biosafety Laboratory, Chinese Academy of Sciences Advanced Customer Cultivation Project (2019ACCP-MS03), the Open Research Fund Program of the State Key Laboratory of Virology of China (2018IOV001).
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JC, GX, and WW conceived and designed the study. Experiments were performed by JC, GZ, MZ, and YL. Data were analyzed by JC and WW. Advice for the project was received from GX and WW. WW and JC wrote the manuscript. All authors read and approved the final manuscript.
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Cao, J., Zhang, G., Zhou, M. et al. Characterizing the Lassa Virus Envelope Glycoprotein Membrane Proximal External Region for Its Role in Fusogenicity. Virol. Sin. 36, 273–280 (2021). https://doi.org/10.1007/s12250-020-00286-3
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DOI: https://doi.org/10.1007/s12250-020-00286-3