Phosphosite Analysis of the Cytomegaloviral mRNA Export Factor pUL69 Reveals Serines with Critical Importance for Recruitment of Cellular Proteins Pin1 and UAP56/URH49.
Cytomegalovirus
/ genetics
DEAD-box RNA Helicases
/ metabolism
HEK293 Cells
Humans
Mutation
NIMA-Interacting Peptidylprolyl Isomerase
/ metabolism
Phosphorylation
RNA, Messenger
/ genetics
RNA, Viral
/ genetics
Serine
/ metabolism
Threonine
/ metabolism
Transcription Factors
/ metabolism
Virus Replication
ICP27
herpesviruses
human cytomegalovirus
mRNA export
pUL69
protein phosphorylation
Journal
Journal of virology
ISSN: 1098-5514
Titre abrégé: J Virol
Pays: United States
ID NLM: 0113724
Informations de publication
Date de publication:
31 03 2020
31 03 2020
Historique:
received:
21
12
2019
accepted:
13
01
2020
pubmed:
24
1
2020
medline:
29
9
2020
entrez:
24
1
2020
Statut:
epublish
Résumé
Human cytomegalovirus (HCMV) encodes the viral mRNA export factor pUL69, which facilitates the cytoplasmic accumulation of mRNA via interaction with the cellular RNA helicase UAP56 or URH49. We reported previously that pUL69 is phosphorylated by cellular CDKs and the viral CDK-like kinase pUL97. Here, we set out to identify phosphorylation sites within pUL69 and to characterize their importance. Mass spectrometry-based phosphosite mapping of pUL69 identified 10 serine/threonine residues as phosphoacceptors. Surprisingly, only a few of these sites localized to the N terminus of pUL69, which could be due to the presence of additional posttranslational modifications, like arginine methylation. As an alternative approach, pUL69 mutants with substitutions of putative phosphosites were analyzed by Phos-tag SDS-PAGE. This demonstrated that serines S46 and S49 serve as targets for phosphorylation by pUL97. Furthermore, we provide evidence that phosphorylation of these serines mediates
Identifiants
pubmed: 31969433
pii: JVI.02151-19
doi: 10.1128/JVI.02151-19
pmc: PMC7108844
pii:
doi:
Substances chimiques
NIMA-Interacting Peptidylprolyl Isomerase
0
RNA, Messenger
0
RNA, Viral
0
Transcription Factors
0
Threonine
2ZD004190S
Serine
452VLY9402
DDX39A protein, human
EC 3.6.1.-
DDX39B protein, human
EC 3.6.1.-
DEAD-box RNA Helicases
EC 3.6.4.13
PIN1 protein, human
EC 5.2.1.8
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2020 American Society for Microbiology.
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