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Interplay between SpaO variants shapes the architecture of the Salmonella type III secretion sorting platform
Open accessPreprint — published as 10.1128/mbio.00155-26 · Paper page
Abstract
Salmonella enterica utilizes a virulence-associated type III secretion system (T3SS) to inject bacterial effectors directly into host cells. Central to this machinery is the sorting platform (SP), a cytosolic assembly whose core scaffolding protein, SpaO, is produced in two isoforms: a full-length (SpaO L ) and a shorter variant (SpaO short ) comprising the C-terminal 101 residues of SpaO L . Although SpaO short is evolutionarily conserved across type III secretion systems, its precise function has remained elusive. Here, we combined a sensitive, real-time translocation assay with site-directed photo-crosslinking to elucidate the role of SpaO short in Salmonella SPI-1 T3SS. We found that while SpaO short is not absolutely required for effector secretion, its absence significantly dampens T3SS-mediated protein delivery. Further biochemical and structural probing revealed that SpaO short is a structural component of the sorting platform, arranged as a homodimer associated to SpaO L via an N-terminal “docking motif.” This interaction occurs while SpaO L is associated with other SP components, supporting a model in which SpaO short is integrated into the SP pods alongside SpaO L , OrgA, and OrgB. Collectively, these findings show that SpaO short , while not strictly essential, functions as a critical structural component of the sorting platform, providing new insights into how Salmonella and related bacteria assemble and maintain these specialized protein-injection systems.
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Highly accurate protein structure prediction with AlphaFold
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Metadata
- Authors
- Soto, J. E., Wang, T., Galán, J. E., & Lara-Tejero, M.
- Affiliations
- Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven CT06536
- Yale University (US)
- Venue
- mBio
- Publisher
- openRxiv
- Published
- 2025-05-15
- Volume
- 17
- Issue
- 4
- Pages
- e0015526-e0015526
- ISSN
- 2150-7511 (ISSN-L)
- PMC
- PMC13059759
- OpenAlex
- W7131884922
- Cited by
- 4
- References
- 1
- Citations by year
- 2026: 4
- FWCI
- 17.58
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- mBio (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (submittedVersion, cc-by)
- PubMed Central (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- University of Missouri
- National Institutes of Health (1r01ai189569-01)
- National Institute of Allergy and Infectious Diseases (R37AI030492)
- All of Us Research Program (R01AI189569, R37AI030492)
- Topics
- Life Sciences › Immunology and Microbiology › Microbiology › Escherichia coli research studies
- Life Sciences › Agricultural and Biological Sciences › Food Science › Salmonella and Campylobacter epidemiology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Salmonella enterica, type III secretion system, injectisome, SPI-1, effector translocation, antivirulence
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