TY - GEN A1 - Zurnic, Irena A1 - Hütter, Sylvia A1 - Rzeha, Ute A1 - Stanke, Nicole A1 - Reh, Juliane A1 - Müllers, Erik A1 - Hamann, Martin V. A1 - Kern, Tobias A1 - Gerresheim, Gesche K. A1 - Lindel, Fabian A1 - Serrao, Erik A1 - Lesbats, Paul A1 - Engelman, Alan N. A1 - Cherepanov, Peter A1 - Lindemann, Dirk T1 - Interactions of prototype foamy virus capsids with host cell polo-like kinases are important for efficient viral DNA integration T2 - Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe N2 - 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. 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. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 580 KW - core protein KW - HIV-1 infection KW - retroviral integration KW - reverse transcription KW - nuclear-localization KW - box domain KW - in-vivo KW - Gag KW - PLK1 KW - phosphorylation Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-411317 SN - 1866-8372 IS - 580 ER -