Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry
Zika virus (ZIKV), a family member in the Flavivirus genus, has re-emerged as a global public health concern. The envelope (E) proteins of flaviviruses play a dual role in viral assembly and entry. To identify the key residues of E in virus entry, we generated a ZIKV trans-complemented particle (ZIK...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2022-12-01
|
| Series: | Emerging Microbes and Infections |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2022.2082888 |
| _version_ | 1818202631961575424 |
|---|---|
| author | Xiao Ma Zhenghong Yuan Zhigang Yi |
| author_facet | Xiao Ma Zhenghong Yuan Zhigang Yi |
| author_sort | Xiao Ma |
| collection | DOAJ |
| description | Zika virus (ZIKV), a family member in the Flavivirus genus, has re-emerged as a global public health concern. The envelope (E) proteins of flaviviruses play a dual role in viral assembly and entry. To identify the key residues of E in virus entry, we generated a ZIKV trans-complemented particle (ZIKVTCP) system, in which a subgenomic reporter replicon was packaged by trans-complementation with expression of CprME. We performed mutagenesis studies of the loop regions that protrude from the surface of the virion in the E ectodomains (DI, DII, DIII). Most mutated ZIKVTCPs exhibited deficient egress. Mutations in DII and in the hinge region of DI and DIII affected prM expression. With a bioorthogonal system, photocrosslinking experiments identified crosslinked intracellular E trimers and demonstrated that egress-deficient mutants in DIII impaired E trimerization. Of these mutants, an E-trimerization-dead mutation D389A that nears the E-E interface between two neighbouring spikes in the immature virion completely abolished viral egress. Several mutations abolished ZIKVTCPs’ entry, without severely affecting viral egress. Further virus binding experiments demonstrated a deficiency of the mutated ZIKVTCPs in virus attachment. Strikingly, synthesized peptide containing residues of two mutants (268-273aa in DII) could bind to host cells and significantly compete for viral attachment and interfere with viral infection, suggesting an important role of these resides in virus entry. Our findings uncovered the requirement for DIII mediated-E trimerization in viral egress, and discovered a key residue group in DII that participates in virus entry. |
| first_indexed | 2024-12-12T03:12:32Z |
| format | Article |
| id | doaj.art-bec9664c19c74051af21269a975c22b7 |
| institution | Directory Open Access Journal |
| issn | 2222-1751 |
| language | English |
| last_indexed | 2024-12-12T03:12:32Z |
| publishDate | 2022-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj.art-bec9664c19c74051af21269a975c22b72022-12-22T00:40:23ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512022-12-011111604162010.1080/22221751.2022.2082888Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entryXiao Ma0Zhenghong Yuan1Zhigang Yi2Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, and Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, People’s Republic of ChinaKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, and Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, People’s Republic of ChinaKey Laboratory of Medical Molecular Virology (MOE/NHC/CAMS), School of Basic Medical Sciences, and Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai, People’s Republic of ChinaZika virus (ZIKV), a family member in the Flavivirus genus, has re-emerged as a global public health concern. The envelope (E) proteins of flaviviruses play a dual role in viral assembly and entry. To identify the key residues of E in virus entry, we generated a ZIKV trans-complemented particle (ZIKVTCP) system, in which a subgenomic reporter replicon was packaged by trans-complementation with expression of CprME. We performed mutagenesis studies of the loop regions that protrude from the surface of the virion in the E ectodomains (DI, DII, DIII). Most mutated ZIKVTCPs exhibited deficient egress. Mutations in DII and in the hinge region of DI and DIII affected prM expression. With a bioorthogonal system, photocrosslinking experiments identified crosslinked intracellular E trimers and demonstrated that egress-deficient mutants in DIII impaired E trimerization. Of these mutants, an E-trimerization-dead mutation D389A that nears the E-E interface between two neighbouring spikes in the immature virion completely abolished viral egress. Several mutations abolished ZIKVTCPs’ entry, without severely affecting viral egress. Further virus binding experiments demonstrated a deficiency of the mutated ZIKVTCPs in virus attachment. Strikingly, synthesized peptide containing residues of two mutants (268-273aa in DII) could bind to host cells and significantly compete for viral attachment and interfere with viral infection, suggesting an important role of these resides in virus entry. Our findings uncovered the requirement for DIII mediated-E trimerization in viral egress, and discovered a key residue group in DII that participates in virus entry.https://www.tandfonline.com/doi/10.1080/22221751.2022.2082888Zika virus (ZIKV)flavivirusenvelope proteinglycoproteinviral egressviral entry |
| spellingShingle | Xiao Ma Zhenghong Yuan Zhigang Yi Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry Emerging Microbes and Infections Zika virus (ZIKV) flavivirus envelope protein glycoprotein viral egress viral entry |
| title | Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry |
| title_full | Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry |
| title_fullStr | Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry |
| title_full_unstemmed | Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry |
| title_short | Identification and characterization of key residues in Zika virus envelope protein for virus assembly and entry |
| title_sort | identification and characterization of key residues in zika virus envelope protein for virus assembly and entry |
| topic | Zika virus (ZIKV) flavivirus envelope protein glycoprotein viral egress viral entry |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2022.2082888 |
| work_keys_str_mv | AT xiaoma identificationandcharacterizationofkeyresiduesinzikavirusenvelopeproteinforvirusassemblyandentry AT zhenghongyuan identificationandcharacterizationofkeyresiduesinzikavirusenvelopeproteinforvirusassemblyandentry AT zhigangyi identificationandcharacterizationofkeyresiduesinzikavirusenvelopeproteinforvirusassemblyandentry |