PXD036455

PXD036455 is an original dataset announced via ProteomeXchange.

Title	Proteome after 3MB-PP1 treatment
Description	The human cytomegalovirus (HCMV) is a member of the beta-herpesvirus family and inflicts life-long latent infections in its hosts. HCMV has been shown to manipulate and dysregulate many cellular processes. On major interactor with the cellular host is the viral kinase pUL97. The UL97 gene is essential for viral replication and kinase-deficient mutants of pUL97 display a severe replication defect. Recently, another group established an analog-sensitive version of the pUL97 protein. This mutant kinase can be treated with a non-hydrolysable ATP analog, thereby inhibiting its kinase function. This process is reversible by removing the ATP analog by media change. We introduced this mutant version of the pUL97 protein into the laboratory strain Ad169 of HCMV, BADwt, creating a BAD-UL97-as1 viral mutant. This mutant virus replicated normally in infected cells in the absence of the ATP analog and maintained its ability to phosphorylate its cellular substrates. However, when treated with the ATP analog, BAD-UL97-as1 displayed a defect in the production of intra- and extracellular viral DNA and in the production of viral progeny. Furthermore, in the presence of 3MB-PP1, the well-established substrate of pUL97, the retinoblastoma protein (Rb) was no longer phosphorylated. This effect was detectable as early as 4 hours post treatment, which allows for studies on pUL97 without the complication of low viral titers. Nevertheless, we observed off-target effects of 3MB-PP1 on several cellular processes, which should be considered with this approach.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:42:59.928.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Uxia Gestal Mato
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-09-02 05:12:38	ID requested
1	2023-03-11 05:46:20	announced
⏵ 2	2023-11-14 08:43:02	announced	2023-11-14: Updated project metadata.

Kr, ä, mer N, Sch, ü, tz M, Mato UG, Herhaus L, Marschall M, Zimmermann C, Recombinant Human Cytomegalovirus Expressing an Analog-Sensitive Kinase pUL97 as Novel Tool for Functional Analyses. Viruses, 14(10):(2022) [pubmed]

submitter keyword: Human, TMT

Lina Herhaus
contact affiliation	Institute of Biochemistry II (IBC2), Johann Wolfgang Goethe-University Hospital, Frankfurt, Germany
contact email	herhaus@em.uni-frankfurt.de
lab head
Uxia Gestal Mato
contact affiliation	Institut für Biochemie II - Goethe Universität Frankfurt
contact email	gestalmato@em.uni-frankfurt.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD036455
     1. Label: PRIDE project
     2. Name: Proteome after 3MB-PP1 treatment