title: The lysine methyltransferase SMYD3 interacts with hepatitis C virus NS5A and is a negative regulator of viral particle production.
abstract: Hepatitis C virus (HCV) is a considerable global health and economic burden. The HCV nonstructural protein (NS) 5A is essential for the viral life cycle. The ability of NS5A to interact with different host and viral proteins allow it to manipulate cellular pathways and regulate viral processes, including RNA replication and virus particle assembly. As part of a proteomic screen, we identified several NS5A-binding proteins, including the lysine methyltransferase SET and MYND domain containing protein 3 (SMYD3). We confirmed the interaction in the context of viral replication by co-immunoprecipitation and co-localization studies. Mutational analyses revealed that the MYND-domain of SMYD3 and domain III of NS5A are required for the interaction. Overexpression of SMYD3 resulted in decreased intracellular and extracellular virus titers, whilst viral RNA replication remained unchanged, suggesting that SMYD3 negatively affects HCV particle production in a NS5A-dependent manner.
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria.
authors.forename: Carol-Ann
authors.initials: CA
authors.lastname: Eberle
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany.
authors.forename: Margarita
authors.initials: M
authors.lastname: Zayas
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria.
authors.forename: Alexey
authors.initials: A
authors.lastname: Stukalov
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria; Max-Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Pichlmair
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany; Department of Molecular Medicine, Via Gabelli 63, 35121 Padua, Italy.
authors.forename: Gualtiero
authors.initials: G
authors.lastname: Alvisi
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria.
authors.forename: André C
authors.initials: AC
authors.lastname: Müller
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria.
authors.forename: Keiryn L
authors.initials: KL
authors.lastname: Bennett
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany. Electronic address: Ralf.Bartenschlager@med.uni-heidelberg.de.
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Lazarettgasse 14, AKH BT 25.3, 1090 Vienna, Austria. Electronic address: gsuperti@cemm.oeaw.ac.at.
authors.forename: Giulio
authors.initials: G
authors.lastname: Superti-Furga
mesh_terms: D002460:Cell Line; D004252:DNA Mutational Analysis; D016174:Hepacivirus / Q000502:physiology*; D011495:Histone-Lysine N-Methyltransferase / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014771:Virion / Q000378:metabolism*; D019065:Virus Assembly; D012324:RNA-Dependent RNA Polymerase
chemical_list: D011495:Histone-Lysine N-Methyltransferase; D017361:Viral Nonstructural Proteins; C506278:SMYD3 protein, human; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: HCV; NS5A; SMYD3; TAP-MS; Virus particle assembly
references.citation: J Biol Chem. 2002 Feb 15;277(7):4906-10
references.pmid: 11733528
references.citation: Virology. 2007 Jul 20;364(1):1-9
references.pmid: 17400273
references.citation: J Virol. 2010 Jun;84(11):5775-89
references.pmid: 20335258
references.citation: J Virol. 2005 Mar;79(5):3187-94
references.pmid: 15709040
references.citation: J Biol Chem. 2004 Nov 19;279(47):48576-87
references.pmid: 15339921
references.citation: Science. 2005 Jul 22;309(5734):623-6
references.pmid: 15947137
references.citation: Trends Microbiol. 2011 Feb;19(2):95-103
references.pmid: 21146993
references.citation: PLoS One. 2011;6(7):e22290
references.pmid: 21779408
references.citation: J Virol. 2006 Nov;80(22):11074-81
references.pmid: 16956946
references.citation: Mol Cell Proteomics. 2014 Jan;13(1):184-203
references.pmid: 24169621
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834
references.citation: J Biol Chem. 2002 Mar 8;277(10):8130-9
references.pmid: 11744739
references.citation: Nucleic Acids Res. 2011 May;39(10):4438-49
references.pmid: 21266482
references.citation: J Gen Virol. 2009 Jun;90(Pt 6):1329-1334
references.pmid: 19264615
references.citation: Med Oncol. 2011 Dec;28 Suppl 1:S91-8
references.pmid: 20957523
references.citation: J Virol. 2005 Jan;79(1):380-92
references.pmid: 15596831
references.citation: J Virol. 2008 Feb;82(3):1073-83
references.pmid: 18032500
references.citation: J Virol. 2007 Dec;81(23):13168-79
references.pmid: 17881454
references.citation: J Virol. 2001 May;75(10):4614-24
references.pmid: 11312331
references.citation: PLoS Pathog. 2008 Mar 28;4(3):e1000035
references.pmid: 18369481
references.citation: Cancer Res. 2012 Feb 1;72(3):810-20
references.pmid: 22194464
references.citation: Cell Host Microbe. 2011 Jan 20;9(1):32-45
references.pmid: 21238945
references.citation: Nat Cell Biol. 2004 Aug;6(8):731-40
references.pmid: 15235609
references.citation: J Clin Gastroenterol. 2011 Feb;45(2):e17-24
references.pmid: 20628308
references.citation: World J Gastroenterol. 2007 Nov 21;13(43):5718-24
references.pmid: 17963297
references.citation: Nat Methods. 2013 Aug;10(8):730-6
references.pmid: 23921808
references.citation: J Virol. 2002 Dec;76(24):13001-14
references.pmid: 12438626
references.citation: Proc Natl Acad Sci U S A. 2006 May 9;103(19):7408-13
references.pmid: 16651538
references.citation: J Biol Chem. 2004 Sep 24;279(39):40835-43
references.pmid: 15247283
references.citation: J Virol. 1995 Jul;69(7):3980-6
references.pmid: 7769656
references.citation: J Mol Biol. 2011 Feb 11;406(1):149-59
references.pmid: 21167177
references.citation: J Virol. 2008 Aug;82(16):7964-76
references.pmid: 18524832
references.citation: Biochem Biophys Res Commun. 2005 Oct 21;336(2):572-8
references.pmid: 16139795
references.citation: Nucleic Acids Res. 1990 Nov 11;18(21):6464
references.pmid: 2173834
references.citation: Drugs Future. 2011 Sep;36(9):691-711
references.pmid: 23378700
references.citation: J Gen Virol. 2003 Mar;84(Pt 3):555-560
references.pmid: 12604805
references.citation: J Proteome Res. 2014 Feb 7;13(2):1147-55
references.pmid: 24400740
references.citation: PLoS Pathog. 2008 Mar 21;4(3):e1000032
references.pmid: 18369478
references.citation: J Virol. 2007 Jan;81(2):588-98
references.pmid: 17079281
references.citation: Nature. 2012 Jul 26;487(7408):486-90
references.pmid: 22810585
references.citation: Mol Syst Biol. 2008;4:230
references.pmid: 18985028
references.citation: Hepatology. 2013 Apr;57(4):1333-42
references.pmid: 23172780
references.citation: Proteomics. 2012 Feb;12(4-5):564-86
references.pmid: 22246820
references.citation: Cancer Res. 2007 Nov 15;67(22):10759-65
references.pmid: 18006819
references.citation: Nat Methods. 2011 Jan;8(1):70-3
references.pmid: 21131968
references.citation: J Virol. 2003 May;77(9):5487-92
references.pmid: 12692249