PXD067302-1

PXD067302 is an original dataset announced via ProteomeXchange.

Title	Dynamics of the CD9 interactome during bacterial infection of epithelial cells by proximity labelling proteomics
Description	Epithelial colonisation is often a critical first step in bacterial pathogenesis, however, different bacterial species utilise several different receptors at the cell membrane interface to adhere to cells. We have previously demonstrated that interference of the human tetraspanin, CD9, can reduce adherence of multiple species of bacteria to epithelial cells by approximately 50%. However, CD9 does not act as a receptor and is responsible for organising and clustering partner proteins commandeered by bacteria for efficient adherence. CD9 can organise numerous host proteins at the cell membrane but the full interactome has not been delineated. Using a novel CD9 proximity-labelling model, we describe a diverse CD9 interactome with 1,837 significantly enriched proteins over four hours. These putative proximal proteins are associated with various cellular pathways including cell adhesion, ECM-receptor interactions, endocytosis, SNARE interactions and adherens and tight junctions. Significant and known interactors of CD9 were enriched including β1 integrins and major immunoglobulin superfamily members but also included several known bacterial adherence receptors including CD44, CD46 and CD147. We further demonstrate dynamism of the interactome during infection at three separate time points with two different bacterial species, Neisseria meningitidis and Staphylococcus aureus. During meningococcal infection, we observed 13 unique significantly enriched proximal proteins associated with CD9 across four hours compared to uninfected cells. However, upon staphylococcal far fewer enriched proximal proteins were identified demonstrating that different bacteria require different host factors during CD9-mediated bacterial adherence. Transient knockdown of CD44 and CD147, candidate receptor proteins identified in our screen, significantly reduced staphylococcal and meningococcal adherence respectively. This effect was ablated in the absence of CD9 or if epithelial cells were treated with a CD9-derived peptide demonstrating the association of these proteins during staphylococcal and meningococcal adherence. We demonstrate for the first time the CD9 interactome of epithelial cells and that bacteria hijack these interactions to efficiently adhere to epithelial cells. This process is bacterial species specific, recruiting a number of different proteins during infection but a host-derived peptide is able to interfere with this process. We have therefore developed a tool that can measure changes within the CD9 interactome after cellular challenge, established a mechanism in which CD9 is used as a universal organiser of bacterial adhesion platforms and demonstrated that this process can be stopped using a CD9-derived peptide.
HostingRepository	PRIDE
AnnounceDate	2025-10-28
AnnouncementXML	Submission_2025-10-28_06:39:47.607.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mark Collins
SpeciesList	scientific name: Neisseria meningitidis serogroup B; NCBI TaxID: NEWT:491; scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606; scientific name: Staphylococcus aureus; NCBI TaxID: NEWT:1280;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-08-13 08:10:12	ID requested
⏵ 1	2025-10-28 06:39:48	announced

10.1111/febs.70291;

Wolverson PA, Fernandes Parreira I, Thompson RH, Collins MO, Shaw JG, Green LR, Dynamics of the CD9 interactome during bacterial infection of epithelial cells by proximity labelling proteomics. FEBS J, ():(2025) [pubmed]

submitter keyword: Neisseria meningitidis, Staphylococcus aureus,CDE9

Mark Collins
contact affiliation	​School of Biosciences University of Sheffield Sheffield, S10 2TN United Kingdom
contact email	mark.collins@sheffield.ac.uk
lab head
Mark Collins
contact affiliation	University of Sheffield
contact email	mark.collins@sheffield.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD067302
     1. Label: PRIDE project
     2. Name: Dynamics of the CD9 interactome during bacterial infection of epithelial cells by proximity labelling proteomics