Human cytomegalovirus pUL97 kinase induces global changes in the infected cell phosphoproteome.

TitleHuman cytomegalovirus pUL97 kinase induces global changes in the infected cell phosphoproteome.
Publication TypeJournal Article
Year of Publication2015
AuthorsOberstein, A, Perlman, DH, Shenk, T, Terry, LJ
JournalProteomics
Volume15
Issue12
Pagination2006-22
Date Published2015 Jun
ISSN1615-9861
KeywordsBlotting, Western, Cells, Cultured, Chromatography, Liquid, Cytomegalovirus, Cytomegalovirus Infections, Fibroblasts, Humans, Phosphoproteins, Phosphorylation, Phosphotransferases (Alcohol Group Acceptor), Protein Interaction Domains and Motifs, Proteome, Proteomics, Tandem Mass Spectrometry, Virus Replication
Abstract

<p>Replication of human cytomegalovirus (HCMV) is regulated in part by cellular kinases and the single viral Ser/Thr kinase, pUL97. The virus-coded kinase augments the replication of HCMV by enabling nuclear egress and altering cell cycle progression. These roles are accomplished through direct phosphorylation of nuclear lamins and the retinoblastoma protein, respectively. In an effort to identify additional pUL97 substrates, we analyzed the phosphoproteome of SILAC-labeled human fibroblasts during infection with either wild-type HCMV or a pUL97 kinase-dead mutant virus. Phosphopeptides were enriched over a titanium dioxide matrix and analyzed by high-resolution MS. We identified 157 unambiguous phosphosites from 106 cellular and 17 viral proteins whose phosphorylation required UL97. Analysis of peptides containing these sites allowed the identification of several candidate pUL97 phosphorylation motifs, including a completely novel phosphorylation motif, LxSP. Substrates harboring the LxSP motif were enriched in nucleocytoplasmic transport functions, including a number of components of the nuclear pore complex. These results extend the known functions of pUL97 and suggest that modulation of nuclear pore function may be important during HCMV replication. </p>

DOI10.1002/pmic.201400607
Alternate JournalProteomics
PubMed ID25867546
PubMed Central IDPMC4522931
Grant ListR56 AI112951 / AI / NIAID NIH HHS / United States
F32AI106175-02 / AI / NIAID NIH HHS / United States
AI112951 / AI / NIAID NIH HHS / United States
F32 AI106175 / AI / NIAID NIH HHS / United States
R01 AI112951 / AI / NIAID NIH HHS / United States