The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms
Paramyxovirus membrane fusion requires an attachment protein for receptor binding and a fusion protein for membrane fusion triggering. Nipah virus (NiV) attachment protein (G) binds to ephrinB2 or -B3 receptors, and fusion protein (F) mediates membrane fusion. NiV-F is a class I fusion protein and i...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2025-01-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/97017 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846093722550272000 |
|---|---|
| author | Qian Wang Jinxin Liu Yuhang Luo Vicky Kliemke Giuliana Leonarda Matta Jingjing Wang Qian Liu |
| author_facet | Qian Wang Jinxin Liu Yuhang Luo Vicky Kliemke Giuliana Leonarda Matta Jingjing Wang Qian Liu |
| author_sort | Qian Wang |
| collection | DOAJ |
| description | Paramyxovirus membrane fusion requires an attachment protein for receptor binding and a fusion protein for membrane fusion triggering. Nipah virus (NiV) attachment protein (G) binds to ephrinB2 or -B3 receptors, and fusion protein (F) mediates membrane fusion. NiV-F is a class I fusion protein and is activated by endosomal cleavage. The crystal structure of a soluble GCN4-decorated NiV-F shows a hexamer-of-trimer assembly. Here, we used single-molecule localization microscopy to quantify the NiV-F distribution and organization on cell and virus-like particle membranes at a nanometer precision. We found that NiV-F on biological membranes forms distinctive clusters that are independent of endosomal cleavage or expression levels. The sequestration of NiV-F into dense clusters favors membrane fusion triggering. The nano-distribution and organization of NiV-F are susceptible to mutations at the hexamer-of-trimer interface, and the putative oligomerization motif on the transmembrane domain. We also show that NiV-F nanoclusters are maintained by NiV-F–AP-2 interactions and the clathrin coat assembly. We propose that the organization of NiV-F into nanoclusters facilitates membrane fusion triggering by a mixed population of NiV-F molecules with varied degrees of cleavage and opportunities for interacting with the NiV-G/receptor complex. These observations provide insights into the in situ organization and activation mechanisms of the NiV fusion machinery. |
| format | Article |
| id | doaj-art-9202a6206d364da6ad936455ff817f4e |
| institution | Kabale University |
| issn | 2050-084X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj-art-9202a6206d364da6ad936455ff817f4e2025-01-02T16:34:21ZengeLife Sciences Publications LtdeLife2050-084X2025-01-011310.7554/eLife.97017The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanismsQian Wang0Jinxin Liu1Yuhang Luo2Vicky Kliemke3Giuliana Leonarda Matta4Jingjing Wang5Qian Liu6https://orcid.org/0000-0002-6174-8982Institute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, CanadaInstitute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, Canada; Mark Wainberg Center for Viral Diseases, Lady Davis Institute, Montreal, CanadaParamyxovirus membrane fusion requires an attachment protein for receptor binding and a fusion protein for membrane fusion triggering. Nipah virus (NiV) attachment protein (G) binds to ephrinB2 or -B3 receptors, and fusion protein (F) mediates membrane fusion. NiV-F is a class I fusion protein and is activated by endosomal cleavage. The crystal structure of a soluble GCN4-decorated NiV-F shows a hexamer-of-trimer assembly. Here, we used single-molecule localization microscopy to quantify the NiV-F distribution and organization on cell and virus-like particle membranes at a nanometer precision. We found that NiV-F on biological membranes forms distinctive clusters that are independent of endosomal cleavage or expression levels. The sequestration of NiV-F into dense clusters favors membrane fusion triggering. The nano-distribution and organization of NiV-F are susceptible to mutations at the hexamer-of-trimer interface, and the putative oligomerization motif on the transmembrane domain. We also show that NiV-F nanoclusters are maintained by NiV-F–AP-2 interactions and the clathrin coat assembly. We propose that the organization of NiV-F into nanoclusters facilitates membrane fusion triggering by a mixed population of NiV-F molecules with varied degrees of cleavage and opportunities for interacting with the NiV-G/receptor complex. These observations provide insights into the in situ organization and activation mechanisms of the NiV fusion machinery.https://elifesciences.org/articles/97017membrane fusionsingle-molecule localization microscopyparamyxovirus |
| spellingShingle | Qian Wang Jinxin Liu Yuhang Luo Vicky Kliemke Giuliana Leonarda Matta Jingjing Wang Qian Liu The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms eLife membrane fusion single-molecule localization microscopy paramyxovirus |
| title | The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms |
| title_full | The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms |
| title_fullStr | The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms |
| title_full_unstemmed | The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms |
| title_short | The nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms |
| title_sort | nanoscale organization of the nipah virus fusion protein informs new membrane fusion mechanisms |
| topic | membrane fusion single-molecule localization microscopy paramyxovirus |
| url | https://elifesciences.org/articles/97017 |
| work_keys_str_mv | AT qianwang thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT jinxinliu thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT yuhangluo thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT vickykliemke thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT giulianaleonardamatta thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT jingjingwang thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT qianliu thenanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT qianwang nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT jinxinliu nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT yuhangluo nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT vickykliemke nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT giulianaleonardamatta nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT jingjingwang nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms AT qianliu nanoscaleorganizationofthenipahvirusfusionproteininformsnewmembranefusionmechanisms |