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Volume 36 Issue 2
April 2021
    Citation: Junyuan Cao, Guangshun Zhang, Minmin Zhou, Yang Liu, Gengfu Xiao, Wei Wang. Characterizing the Lassa Virus Envelope Glycoprotein Membrane Proximal External Region for Its Role in Fusogenicity .VIROLOGICA SINICA, 2021, 36(2) : 273-280.  http://dx.doi.org/10.1007/s12250-020-00286-3
    shu

    Characterizing the Lassa Virus Envelope Glycoprotein Membrane Proximal External Region for Its Role in Fusogenicity

    • 1.

      State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China

    • 2.

      University of the Chinese Academy of Sciences, Beijing 100049, China

    • 3.

      College of Life Sciences, Nankai University, Tianjin 300353, China

    • Corresponding author: Gengfu Xiao, xiaogf@wh.iov.cn, ORCID: 0000-0001-9401-235X
      Wei Wang, wangwei@wh.iov.cn, ORCID: 0000-0002-8696-9773
    • Received Date: 30 April 2020
      Accepted Date: 22 July 2020
      Published Date: 08 September 2020
      Available online: 01 April 2021

    • 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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      Characterizing the Lassa Virus Envelope Glycoprotein Membrane Proximal External Region for Its Role in Fusogenicity

        Corresponding author: Gengfu Xiao, xiaogf@wh.iov.cn
        Corresponding author: Wei Wang, wangwei@wh.iov.cn
      • 1. State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China
      • 2. University of the Chinese Academy of Sciences, Beijing 100049, China
      • 3. College of Life Sciences, Nankai University, Tianjin 300353, China
      • Received Date: 30 April 2020
      • Accepted Date: 22 July 2020
      • Published Date: 08 September 2020

      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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