PXD030816-2

PXD030816 is an original dataset announced via ProteomeXchange.

Title	The ubiquitin landscape of the Influenza A Virus polymerase
Description	During influenza A virus (IAV) infections, viral proteins are targeted by cellular E3 ligases for modification with ubiquitin. Here, we decipher and functionally explore the ubiquitin landscape of the IAV polymerase during infection of human alveolar epithelial cells by applying mass spectrometry analysis of immuno-purified K-ε-GG- (di-glycyl)-remnant-bearing peptides. We identified 59 modified lysines across all three subunits of the viral polymerase of which 17 distinctively affected mRNA transcription, vRNA replication and the generation of recombinant viruses via non-proteolytic mechanisms. Moreover, our results demonstrate that the ubiquitinated residue K578 in the PB1 thumb domain is crucial for the dynamic structural transitions of the viral polymerase that are required for vRNA replication. Mutations K578A and K578R impeded the steps of cRNA stabilization and vRNA transcription, respectively, and affected NP binding as well as polymerase dimerization. Collectively, our results indicate that ubiquitin-mediated disruption of the charge-dependent interaction between PB1-K578 and PB2-E72 is required to coordinate polymerase dimerization and facilitate vRNA replication, which demonstrates that IAV exploit the cellular ubiquitin system to modulate the activity of the viral polymerase for the regulation of viral replication.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:41:52.506.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD030816
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Hannes Drexler
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Influenza A virus (A/Wilson-Smith/1933(H1N1)); NCBI TaxID: 381518;
ModificationList	ubiquitinylated lysine; ubiquitination signature dipeptidyl lysine; iodoacetamide derivatized residue
Instrument	Q Exactive HF; LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-07 12:42:51	ID requested
1	2023-03-11 03:28:40	announced
⏵ 2	2023-11-14 08:41:53	announced	2023-11-14: Updated project metadata.

10.6019/PXD030816;

G, ü, nl F, Krischuns T, Schreiber JA, Henschel L, Wahrenburg M, Drexler HCA, Leidel SA, Cojocaru V, Seebohm G, Mellmann A, Schwemmle M, Ludwig S, Brunotte L, The ubiquitination landscape of the influenza A virus polymerase. Nat Commun, 14(1):787(2023) [pubmed]

submitter keyword: Influenza A virus, PB1, affinity enrichment, PA, viral RNA-dependent RNA polymerase, ubiquitination, K-ε-GG motif,LC-MSMS, PB2

Hannes C. A. Drexler
contact affiliation	Max Planck Institute for Molecular Biomedicine Bioanalytical Mass Spectrometry Röntgenstr. 20 48149 Münster Germany
contact email	hannes.drexler@mpi-muenster.mpg.de
lab head
Hannes Drexler
contact affiliation	Bioanalytical Mass Spectrometry, Max Planck Insitute for Molecular Biomedicine, Roentgenstr. 20, 48149 Münster, Germany
contact email	hannes.drexler@mpi-muenster.mpg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD030816
     1. Label: PRIDE project
     2. Name: The ubiquitin landscape of the Influenza A Virus polymerase