Presynaptic targeting of botulinum neurotoxin type A requires a tripartite PSG‐Syt1‐SV2 plasma membrane nanocluster for synaptic vesicle entry
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Abstract
The unique nerve terminal targeting of botulinum neurotoxin type A (BoNT/A) is due to its capacity to bind two receptors on the neuronal plasma membrane: polysialoganglioside (PSG) and synaptic vesicle glycoprotein 2 (SV2). Whether and how PSGs and SV2 may coordinate other proteins for BoNT/A recruitment and internalization remains unknown. Here, we demonstrate that the targeted endocytosis of BoNT/A into synaptic vesicles (SVs) requires a tripartite surface nanocluster. Live‐cell super‐resolution imaging and electron microscopy of catalytically inactivated BoNT/A wildtype and receptor‐binding‐deficient mutants in cultured hippocampal neurons demonstrated that BoNT/A must bind coincidentally to a PSG and SV2 to target synaptic vesicles. We reveal that BoNT/A simultaneously interacts with a preassembled PSG‐synaptotagmin‐1 (Syt1) complex and SV2 on the neuronal plasma membrane, facilitating Syt1‐SV2 nanoclustering that controls endocytic sorting of the toxin into synaptic vesicles. Syt1 CRISPRi knockdown suppressed BoNT/A‐ and BoNT/E‐induced neurointoxication as quantified by SNAP‐25 cleavage, suggesting that this tripartite nanocluster may be a unifying entry point for selected botulinum neurotoxins that hijack this for synaptic vesicle targeting.
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Cites
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- Authors
- Joensuu, M., Syed, P., Saber, S. H., Lanoue, V., Wallis, T. P., Rae, J., Blum, A., Gormal, R. S., Small, C., Sanders, S., Jiang, A., Mahrhold, S., Krez, N., Cousin, M. A., Cooper‐White, R., Cooper‐White, J. J., Collins, B. M., Parton, R. G., Balistreri, G., Rummel, A., & Meunier, F. A.
- Affiliations
- The University of Queensland (AU)
- Medizinische Hochschule Hannover (DE)
- Simons Initiative for the Developing Brain (GB)
- University of Helsinki (FI)
- Corresponding author
- Joensuu, Merja
- Meunier, Frédéric A
- ORCID
- Venue
- The EMBO Journal
- Published
- 2023-05-25
- Volume
- 42
- Issue
- 13
- Pages
- e112095-e112095
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC10308369
- OpenAlex
- W4378172982
- Cited by
- 34
- References
- 8
- Citations by year
- 2026: 10 · 2025: 11 · 2024: 9 · 2023: 4
- FWCI
- 5.78
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- The EMBO Journal (publishedVersion, other-oa)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Työväentutkimus Vuosikirja (submittedVersion)
- Edinburgh Research Explorer (University of Edinburgh) (submittedVersion, cc-by)
- Funders
- Academy of Finland (318434)
- Department of Education and Training | Australian Research Council (DE190100565, LE130100078)
- DHAC | National Health and Medical Research Council (1120381, 1155794, 1140064, 1150083, 1156489, 1136021)
- German Federal Ministry of Education and Research (031L0111B, 13N15512)
- The University of Queensland Amplify Fellowship
- Topics
- Health Sciences › Medicine › Neurology › Botulinum Toxin and Related Neurological Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- botulinum toxin type A, botulinum neurotoxin type E, polysialogangliosides, SV2, synaptotagmin-1, synaptic vesicle endocytosis, toxin internalization, SNAP-25 cleavage, hippocampal neurons, receptor binding
- MeSH terms
- Animals, Cell Membrane, Gangliosides, Membrane Glycoproteins, Nerve Tissue Proteins, Synaptic Vesicles, Botulinum Toxins, Type A, Rats
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