PXD056320-1

PXD056320 is an original dataset announced via ProteomeXchange.

Title	To be or not to be phosphorylated: Understanding the role of Ebola virus nucleoprotein in the dynamic interplay with the transcriptional activator VP30 and the host phosphatase PP2A-B56
Description	Ebola virus (EBOV) transcription is essentially regulated via dynamic dephosphorylation of its viral transcription activator VP30 by the host phosphatase PP2A. The nucleoprotein NP has emerged as a third key player in the regulation of this process by recruiting both the regulatory subunit B56 of PP2A and its substrate VP30 to initiate VP30 dephosphorylation and hence viral transcription. Both binding sites are located in close proximity to each other in NP`s C-terminal disordered region. This study investigates NP's role in VP30 dephosphorylation and transcription activation, focusing on the spatial requirements of NP’s binding sites. Increasing the distance between PP2A-B56 and VP30 at the NP interface revealed that close spatial and orientational contact is necessary for efficient VP30 dephosphorylation and viral transcription. Longer distances were lethal for recombinant EBOV except when a compensatory mutation, NP T603I, occurred. This mutation, located between the NP binding sites for PP2A-B56 and VP30, fully restored functionality. Mass spectrometry showed that T603 is phosphorylated in recEBOV-NPwt virions. Mutational analysis indicated that T603I facilitates VP30 dephosphorylation in otherwise lethal recEBOV and that dynamic phosphorylation of NP T603 is important for efficient primary viral transcription in the WT context. These findings emphasize the critical and evolutionarily pressured interplay between VP30 and PP2A-B56 within the NP C-terminal disordered region and highlight the important role of NP on the regulation of viral transcription during the EBOV life cycle.
HostingRepository	PRIDE
AnnounceDate	2025-01-14
AnnouncementXML	Submission_2025-01-14_02:58:34.558.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Uwe Linne
SpeciesList	scientific name: Zaire ebolavirus; NCBI TaxID: NCBITaxon:186538; scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Ebolavirus; NCBI TaxID: 186536;
ModificationList	phosphorylated residue
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2024-09-27 02:36:01	ID requested
⏵ 1	2025-01-14 02:58:34	announced

10.1080/22221751.2024.2447612;

submitter keyword: Ebola virus, host phosphatase PP2A, nucleoprotein, viral transcription, phosphorylation, VP30, regulation

Dr. Uwe Linne
contact affiliation	Philipps-Universität Marburg Faculty of Chemistry Hans-Meerwein-Str. 4 35043 Marburg GERMANY
contact email	linneu@staff.uni-marburg.de
lab head
Uwe Linne
contact affiliation	Head of Core Facility Mass Spectrometry, Faculty of Chemistry, Philipps-University Marburg
contact email	linneu@staff.uni-marburg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD056320
     1. Label: PRIDE project
     2. Name: To be or not to be phosphorylated: Understanding the role of Ebola virus nucleoprotein in the dynamic interplay with the transcriptional activator VP30 and the host phosphatase PP2A-B56