<i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
Asparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are <i>N</i>-glycosylated. The prM protein of all ZIKV strains contains a single <i>...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-08-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/12/9/925 |
| _version_ | 1827708000810303488 |
|---|---|
| author | Yong-Dae Gwon Eva Zusinaite Andres Merits Anna K. Överby Magnus Evander |
| author_facet | Yong-Dae Gwon Eva Zusinaite Andres Merits Anna K. Överby Magnus Evander |
| author_sort | Yong-Dae Gwon |
| collection | DOAJ |
| description | Asparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are <i>N</i>-glycosylated. The prM protein of all ZIKV strains contains a single <i>N</i>-linked glycosylation site, while not all strains contain an N-linked site in the E protein. Our aim was to examine the impact of prM and E N-linked glycosylation on ZIKV infectivity and cell trafficking. Using a ZIKV infectious clone, we found that when the <i>N</i>-glycan sites were removed, the prM- and the prM/E-double mutants did not produce an infectious virus in the supernatant. Further, by using ZIKV prME constructs, we found that <i>N</i>-glycosylation was necessary for effective secretion of ZIKV virions. The absence of the <i>N</i>-glycan on prM or E caused protein aggregation in the rough endoplasmatic reticulum (ER) compartment. The aggregation was more pronounced for the prM-mutation, and the mutant virus lost the ER-Golgi intermediate compartment (ERGIC) localization. In addition, lack of the <i>N</i>-glycan on prM induced nuclear translocation of CCAAT-enhancer-binding protein homologous protein (CHOP), an ER stress marker. To conclude, we show that the prM <i>N</i>-glycan is essential for the ZIKV infectious cycle, and plays an important role in viral protein trafficking, protein folding, and virion assembly<b>.</b> |
| first_indexed | 2024-03-10T16:56:32Z |
| format | Article |
| id | doaj.art-10b2ea36864347b397142cbf94cdd0b2 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T16:56:32Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-10b2ea36864347b397142cbf94cdd0b22023-11-20T11:05:32ZengMDPI AGViruses1999-49152020-08-0112992510.3390/v12090925<i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life CycleYong-Dae Gwon0Eva Zusinaite1Andres Merits2Anna K. Överby3Magnus Evander4Department of Clinical Microbiology, Virology, Umeå University, 901 85 Umeå, SwedenInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaDepartment of Clinical Microbiology, Virology, Umeå University, 901 85 Umeå, SwedenDepartment of Clinical Microbiology, Virology, Umeå University, 901 85 Umeå, SwedenAsparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are <i>N</i>-glycosylated. The prM protein of all ZIKV strains contains a single <i>N</i>-linked glycosylation site, while not all strains contain an N-linked site in the E protein. Our aim was to examine the impact of prM and E N-linked glycosylation on ZIKV infectivity and cell trafficking. Using a ZIKV infectious clone, we found that when the <i>N</i>-glycan sites were removed, the prM- and the prM/E-double mutants did not produce an infectious virus in the supernatant. Further, by using ZIKV prME constructs, we found that <i>N</i>-glycosylation was necessary for effective secretion of ZIKV virions. The absence of the <i>N</i>-glycan on prM or E caused protein aggregation in the rough endoplasmatic reticulum (ER) compartment. The aggregation was more pronounced for the prM-mutation, and the mutant virus lost the ER-Golgi intermediate compartment (ERGIC) localization. In addition, lack of the <i>N</i>-glycan on prM induced nuclear translocation of CCAAT-enhancer-binding protein homologous protein (CHOP), an ER stress marker. To conclude, we show that the prM <i>N</i>-glycan is essential for the ZIKV infectious cycle, and plays an important role in viral protein trafficking, protein folding, and virion assembly<b>.</b>https://www.mdpi.com/1999-4915/12/9/925Zika virus<i>N</i>-glycosylationpre-membraneenvelopevirus life cycle |
| spellingShingle | Yong-Dae Gwon Eva Zusinaite Andres Merits Anna K. Överby Magnus Evander <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle Viruses Zika virus <i>N</i>-glycosylation pre-membrane envelope virus life cycle |
| title | <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle |
| title_full | <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle |
| title_fullStr | <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle |
| title_full_unstemmed | <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle |
| title_short | <i>N</i>-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle |
| title_sort | i n i glycosylation in the pre membrane protein is essential for the zika virus life cycle |
| topic | Zika virus <i>N</i>-glycosylation pre-membrane envelope virus life cycle |
| url | https://www.mdpi.com/1999-4915/12/9/925 |
| work_keys_str_mv | AT yongdaegwon iniglycosylationinthepremembraneproteinisessentialforthezikaviruslifecycle AT evazusinaite iniglycosylationinthepremembraneproteinisessentialforthezikaviruslifecycle AT andresmerits iniglycosylationinthepremembraneproteinisessentialforthezikaviruslifecycle AT annakoverby iniglycosylationinthepremembraneproteinisessentialforthezikaviruslifecycle AT magnusevander iniglycosylationinthepremembraneproteinisessentialforthezikaviruslifecycle |