PXD033391

PXD033391 is an original dataset announced via ProteomeXchange.

Title	Determining kinome changes in response to Cytomegalovirus US28 expression and identifying US28 interacting proteins
Description	Cytomegalovirus (CMV) reactivation of latent infection following immune dysregulation remains a serious risk for patients, often causing significant morbidity and mortality. Herein, we demonstrate the CMV-encoded GPCR, US28, in coordination with cellular Ephrin receptor A2 (EphA2), attenuates MAPK signaling, thereby limiting viral replication in latently-infected primary monocytes. To define the underlying mechanism by which US28 modulates MAPK signaling during latency, we used two unbiased approaches to: 1) profile kinome changes in response to US28 expression, and 2) define US28 interacting partners that could influence its function(s). First, we evaluated changes in the cellular kinome in response to US28 expression in THP-1 monocytic cells compared to control cells using multiplexed kinase inhibitor beads coupled with mass spectrometry (MIB-MS) to quantitatively measure kinase expression and abundance. Next, we leveraged proximity labeling technology, whereby we inserted the biotin ligase gene, birA, conjugated to a C-terminal hemagglutinin (HA) epitope tag in-frame with the US28 ORF to generate TB40/E-mCherry-US28-bioID-HA. We then infected fibroblasts with US28-bioID-HA, added biotin 18 h prior to harvesting the cells at 48 h post-infection (hpi), and analyzed the biotin-conjugated proteins following streptavidin capture by affinity purification-mass spectrometry (AP-MS).
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:42:36.491.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Laura Herring
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-04-23 15:30:58	ID requested
1	2023-03-11 03:58:04	announced
⏵ 2	2023-11-14 08:42:44	announced	2023-11-14: Updated project metadata.

Wass AB, Krishna BA, Herring LE, Gilbert TSK, Nukui M, Groves IJ, Dooley AL, Kulp KH, Matthews SM, Rotroff DM, Graves LM, O'Connor CM, Cytomegalovirus US28 regulates cellular EphA2 to maintain viral latency. Sci Adv, 8(43):eadd1168(2022) [pubmed]

submitter keyword: viral infection,affinity purification-mass spectrometry, kinome

Christine O'Connor
contact affiliation	Genomic Medicine, Infection Biology, Lerner Research Institute, Cleveland Clinic
contact email	oconnoc6@ccf.org
lab head
Laura Herring
contact affiliation	UNC-Chapel Hill
contact email	laura_herring@med.unc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD033391
     1. Label: PRIDE project
     2. Name: Determining kinome changes in response to Cytomegalovirus US28 expression and identifying US28 interacting proteins