title: Interactions of Prototype Foamy Virus Capsids with Host Cell Polo-Like Kinases Are Important for Efficient Viral DNA Integration.
abstract: 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.
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Irena
authors: I
authors: Zurnic
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Sylvia
authors: S
authors: Hütter
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Ute
authors: U
authors: Rzeha
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Nicole
authors: N
authors: Stanke
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Juliane
authors: J
authors: Reh
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Erik
authors: E
authors: Müllers
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Martin V
authors: MV
authors: Hamann
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Tobias
authors: T
authors: Kern
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Gesche K
authors: GK
authors: Gerresheim
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Fabian
authors: 0000-0002-3028-1646
authors: F
authors: Lindel
authors: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, United States of America.
authors: Erik
authors: E
authors: Serrao
authors: Clare Hall Laboratories, The Francis Crick Institute, South Mimms, United Kingdom.
authors: Paul
authors: P
authors: Lesbats
authors: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, United States of America.
authors: Alan N
authors: AN
authors: Engelman
authors: Clare Hall Laboratories, The Francis Crick Institute, South Mimms, United Kingdom.
authors: Peter
authors: P
authors: Cherepanov
authors: Institute of Virology, Medical Faculty "Carl Gustav Carus", Technische Universität Dresden, Dresden, Germany.|CRTD/DFG-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany.
authors: Dirk
authors: D
authors: Lindemann
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D002213:Capsid / Q000378:metabolism*; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010766:Phosphorylation / Q000235:genetics; D000072417:Protein Domains; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D012192:Retroviridae Infections / Q000235:genetics / Q000378:metabolism*; D016092:Spumavirus / Q000235:genetics / Q000378:metabolism*; D016662:Virus Integration / Q000502:physiology*
chemical_list: D015683:Gene Products, gag; C496126:PLK2 protein, human; D017346:Protein Serine-Threonine Kinases; C530504:Plk2 protein, rat; C444640:serum-inducible kinase