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Interactions of prototype foamy virus capsids with host cell polo-like kinases are important for efficient viral DNA integration

  • Unlike for other retroviruses, only a few host cell factors that aid the replication of foamy viruses (FVs) via interaction with viral structural components are known. Using a yeast-two-hybrid (Y2H) screen with prototype FV (PFV) Gag protein as bait we identified human polo-like kinase 2 (hPLK2), a member of cell cycle regulatory kinases, as a new interactor of PFV capsids. Further Y2H studies confirmed interaction of PFV Gag with several PLKs of both human and rat origin. A consensus Ser-Thr/Ser-Pro (S-T/S-P) motif in Gag, which is conserved among primate FVs and phosphorylated in PFV virions, was essential for recognition by PLKs. In the case of rat PLK2, functional kinase and polo-box domains were required for interaction with PFV Gag. Fluorescently-tagged PFV Gag, through its chromatin tethering function, selectively relocalized ectopically expressed eGFP-tagged PLK proteins to mitotic chromosomes in a Gag STP motif-dependent manner, confirming a specific and dominant nature of the Gag-PLK interaction in mammalian cells. TheUnlike for other retroviruses, only a few host cell factors that aid the replication of foamy viruses (FVs) via interaction with viral structural components are known. Using a yeast-two-hybrid (Y2H) screen with prototype FV (PFV) Gag protein as bait we identified human polo-like kinase 2 (hPLK2), a member of cell cycle regulatory kinases, as a new interactor of PFV capsids. Further Y2H studies confirmed interaction of PFV Gag with several PLKs of both human and rat origin. A consensus Ser-Thr/Ser-Pro (S-T/S-P) motif in Gag, which is conserved among primate FVs and phosphorylated in PFV virions, was essential for recognition by PLKs. In the case of rat PLK2, functional kinase and polo-box domains were required for interaction with PFV Gag. Fluorescently-tagged PFV Gag, through its chromatin tethering function, selectively relocalized ectopically expressed eGFP-tagged PLK proteins to mitotic chromosomes in a Gag STP motif-dependent manner, confirming a specific and dominant nature of the Gag-PLK interaction in mammalian cells. The functional relevance of the Gag-PLK interaction was examined in the context of replication-competent FVs and single-round PFV vectors. Although STP motif mutated viruses displayed wild type (wt) particle release, RNA packaging and intra-particle reverse transcription, their replication capacity was decreased 3-fold in single-cycle infections, and up to 20-fold in spreading infections over an extended time period. Strikingly similar defects were observed when cells infected with single-round wt Gag PFV vectors were treated with a pan PLK inhibitor. Analysis of entry kinetics of the mutant viruses indicated a post-fusion defect resulting in delayed and reduced integration, which was accompanied with an enhanced preference to integrate into heterochromatin. We conclude that interaction between PFV Gag and cellular PLK proteins is important for early replication steps of PFV within host cells.show moreshow less

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Metadaten
Author:Irena Zurnic, Sylvia Hütter, Ute Rzeha, Nicole Stanke, Juliane Reh, Erik MüllersORCiD, Martin V. Hamann, Tobias Kern, Gesche K. Gerresheim, Fabian Lindel, Erik Serrao, Paul Lesbats, Alan N. Engelman, Peter CherepanovORCiD, Dirk Lindemann
URN:urn:nbn:de:kobv:517-opus4-411317
DOI:https://doi.org/10.25932/publishup-41131
ISSN:1866-8372
Parent Title (English):Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe
Series (Serial Number):Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe (580)
Document Type:Postprint
Language:English
Date of first Publication:2019/02/07
Year of Completion:2016
Publishing Institution:Universität Potsdam
Release Date:2019/02/07
Tag:Gag; HIV-1 infection; PLK1; box domain; core protein; nuclear-localization; phosphorylation; retroviral integration; reverse transcription
in-vivo
Issue:580
Pagenumber:36
Source:PLoS Pathogens 12 (2016) 8, Art. e1005860 DOI 10.1371/journal.ppat.1005860
Organizational units:Mathematisch-Naturwissenschaftliche Fakultät
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Peer Review:Referiert
Publication Way:Open Access
Grantor:Public Library of Science (PLOS)
Licence (German):License LogoCreative Commons - Namensnennung, 4.0 International