PXD015883

PXD015883 is an original dataset announced via ProteomeXchange.

Title	Proximity Labeling to Map Host-Pathogen Interactions at the Membrane of a Bacteria Containing Vacuole in Chlamydia trachomatis Infected Human Cells
Description	Many intracellular bacteria, including the obligate intracellular pathogen Chlamydia trachomatis, grow within a membrane-bound “bacteria containing vacuole” (BCV). Secreted cytosolic effectors modulate host activity, but an understanding of the host-pathogen interactions that occur at the BCV membrane is limited by the difficulty in purifying membrane fractions from infected host cells. We used the ascorbate peroxidase proximity labeling system (APEX2), which labels proximal proteins with biotin in vivo, to study the protein-protein interactions that occur at the chlamydial vacuolar, or inclusion, membrane. An in vivo understanding of the secreted chlamydial inclusion membrane protein (Inc) interactions (e.g., Inc-Inc and Inc-eukaryotic protein) and how these contribute to overall host-chlamydial interactions at this unique membrane is lacking. We hypothesize some Incs organize the inclusion membrane whereas other Incs bind eukaryotic proteins to promote chlamydial-host interactions. To study this, Incs fused to APEX2 were expressed in C. trachomatis L2. Affinity purification-mass spectrometry (AP-MS) identified biotinylated proteins, which were analyzed for statistical significance using Significance Analysis of INTeractome (SAINT). Broadly supporting both Inc-Inc and Inc-host interactions, our Inc-APEX2 constructs labeled Incs as well as known and previously unreported eukaryotic proteins localizing to the inclusion. We demonstrate that endogenous LRRFIP1 (LRRF1) is recruited to the inclusion by the Inc, CT226, using bacterial two-hybrid and co-immunoprecipitation assays. We further demonstrate interactions between CT226 and the Incs used in our study to reveal a model for inclusion membrane organization. Combined, our data highlight the utility of APEX2 to capture the complex in vivo protein-protein interactions at the chlamydial inclusion.
HostingRepository	PRIDE
AnnounceDate	2019-10-25
AnnouncementXML	Submission_2019-10-25_05:11:06.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD015883
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Macy Olson
SpeciesList	scientific name: Chlamydia trachomatis 434/Bu; NCBI TaxID: 471472;
ModificationList	Biotin-tyramide; Oxidation; Carbamidomethyl
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2019-10-18 01:45:24	ID requested
⏵ 1	2019-10-25 05:11:07	announced

Olson MG, Widner RE, Jorgenson LM, Lawrence A, Lagundzin D, Woods NT, Ouellette SP, Rucks EA, Proximity Labeling To Map Host-Pathogen Interactions at the Membrane of a Bacterium-Containing Vacuole in Chlamydia trachomatis-Infected Human Cells. Infect Immun, 87(11):(2019) [pubmed]

submitter keyword: APEX2, Chlamydia, Inclusion, proximity labeling

Elizabeth A. Rucks and Scot P. Ouellette
contact affiliation	Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.
contact email	lisa.rucks@unmc.edu
lab head
Macy Olson
contact affiliation	University of Nebraska Medical Center
contact email	macy.olson@unmc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD015883
     1. Label: PRIDE project
     2. Name: Proximity Labeling to Map Host-Pathogen Interactions at the Membrane of a Bacteria Containing Vacuole in Chlamydia trachomatis Infected Human Cells