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Gonet, A.

Publications and source records attributed to Gonet, A..

2 recordsLinked to original sources

Visual surfaceomics reveals the substrate-scavenging cell surface of Asgard archaea

The emergence of eukaryotes from Asgard archaeal ancestors likely required a specialized cell envelope that enabled sustained physical and metabolic interactions with proto-mitochondrial symbionts. While the protrusion-rich cell architecture of Asgard archaea may enable such interactions, the molecular composition of their cell surface and its role in establishing cell-cell contacts remains unknown. Here, we developed a "visual surfaceomics" approach, integrating subtomogram averaging, predictive structural modeling, transcriptomics, proteomics, and expansion microscopy to identify surface proteins in situ. We show that Asgard cell bodies and protrusions lack an archaea-typical paracrystalline S-layer and are instead decorated by two main classes of surface proteins: subtilisin-like serine proteases (SubSs) and oligopeptide-binding proteins (OppAs). SubSs were detected as membrane-anchored dimers and were sparsely distributed across the surface. OppAs showed near-total membrane coverage but lacked any rigid, higher order assembly. Thermal shift assays and single particle cryo-EM showed that OppA binds oligopeptides in vitro. Comparative genomics, transcriptomics, and proteomics suggest that this peptidophilic surface architecture is conserved across Asgard archaeal phyla. Finally, we discovered that the cell surface was remodeled at cell-cell contact sites. Altogether, our integrative approach revealed a metabolically specialized and structurally flexible cell surface that couples extracellular protein degradation with peptide capture and uptake, while potentially contributing to establishing and maintaining the symbiotic interactions of Asgard archaea.

microbiology↗

A Seven-Protein Assembly Promotes Stability, Neutralisation and Secretion of the T7SSb LXG-effector TelE

Streptococcus gallolyticus subsp. gallolyticus (SGG) is a gut pathobiont associated with colorectal cancer. Like many Firmicutes, SGG utilizes a specialized Type VII Secretion System (T7SSb) to export WXG100 and LXG proteins, the latter involved in bacterial competition. We previously identified TelE, an LXG protein whose C-terminus mediates membrane pore formation in Escherichia coli. In SGG UCN34, TelE (Gallo_0562) is co-expressed with six proteins (Gallo_0559 to Gallo_0565), including its immunity protein TipE (Gallo_0565). Here we show that the absence of those co-expressed proteins affects TelE stability and secretion. These proteins associate with TelE to form a soluble and stable seven-protein complex. Gallo_0559 and Gallo_0560 interact with the N-terminal LXG domain of TelE, Gallo_0561 binds to its central region, while the six-transmembrane protein Gallo_0563, together with Gallo_0564 and TipE associates with its C-terminal domain. These findings describe a new modular complex that stabilizes TelE while reducing its toxicity and optimizing its T7SSb-mediated delivery.

microbiology↗