PXD004797

PXD004797 is an original dataset announced via ProteomeXchange.

Title	Comparative proteomics of purified pathogen vacuoles correlates intracellular replication of Legionella pneumophila with the small GTPase Rap1
Description	Legionella pneumophila is an opportunistic bacterial pathogen that causes a severe lung infection termed “Legionnaires’ disease”. The pathogen replicates in environmental protozoa as well as in macrophages within a unique membrane-bound compartment, the Legionella-containing-vacuole (LCV). Formation of the pathogen vacuole requires the bacterial Icm/Dot type IV secretion system (T4SS), which translocates ca. 300 effector proteins into host cells, where they subvert pivotal host processes and govern LCV formation. The L. pneumophila “pentuple” mutant lacks 5 gene clusters comprising 12% of the genome and 30% of the effector proteins. The mutant strain replicates in murine bone marrow-derived macrophages but not in Dictyostelium discoideum amoeba. In order to elucidate the host cell proteome defining a replication permissive compartment, we compare here the proteomes of intact LCVs isolated from D. discoideum or murine RAW 264.7 macrophages infected with the L. pneumophila parental strain Lp02 or the “pentuple” mutant. Proteome analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed host proteins specifically localizing to LCVs harbouring strain Lp02 or the “pentuple” mutant, respectively. The small GTPase Rap1 was identified on D. discoideum LCVs containing strain Lp02 but not the “pentuple” mutant and on macrophage LCVs containing either strain. The localization pattern of Rap1 on D. discoideum LCVs was confirmed by fluorescence microscopy, and depletion of Rap1 in A549 epithelial cells by RNA interference significantly reduced the intracellular growth of L. pneumophila. Thus, comparative proteomics identified Rap1 as a novel LCV host component implicated in intracellular replication of L. pneumophila.
HostingRepository	PRIDE
AnnounceDate	2017-02-10
AnnouncementXML	Submission_2017-02-14_03:42:36.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andreas Otto
SpeciesList	scientific name: Legionella; NCBI TaxID: 445;
ModificationList	monohydroxylated residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2016-08-15 02:22:29	ID requested
1	2017-02-10 06:21:04	announced
⏵ 2	2017-02-14 03:42:37	announced	Updated publication reference for PubMed record(s): 28183814.

Schm, ö, lders J, Manske C, Otto A, Hoffmann C, Steiner B, Welin A, Becher D, Hilbi H, with the Small GTPase Ras-related protein 1 (Rap1). Mol Cell Proteomics, 16(4):622-641(2017) [pubmed]

curator keyword: Biomedical

submitter keyword: Amoeba, bacterial effector protein, Dictyostelium discoideum, small GTPase, host-pathogen interactions, Legionella, macrophage, pathogen vacuole, phosphoinositide lipid, proteomics, type IV secretion

Andreas Otto
contact affiliation	Institute of Microbiology, Ernst-Moritz-Arndt University Greifswald, 17489 Greifswald, Germany
contact email	andreas.otto@uni-greifswald.de
lab head
Andreas Otto
contact affiliation	Institute for Microbiology
contact email	andreas.otto@uni-greifswald.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2017/02/PXD004797

PRIDE project URI

• PRIDE
  1. PXD004797
     1. Label: PRIDE project
     2. Name: Comparative proteomics of purified pathogen vacuoles correlates intracellular replication of Legionella pneumophila with the small GTPase Rap1