title: In vitro assembly of virus-like particles of a gammaretrovirus, the murine leukemia virus XMRV.
abstract: Immature retroviral particles are assembled by self-association of the structural polyprotein precursor Gag. During maturation the Gag polyprotein is proteolytically cleaved, yielding mature structural proteins, matrix (MA), capsid (CA), and nucleocapsid (NC), that reassemble into a mature viral particle. Proteolytic cleavage causes the N terminus of CA to fold back to form a β-hairpin, anchored by an internal salt bridge between the N-terminal proline and the inner aspartate. Using an in vitro assembly system of capsid-nucleocapsid protein (CANC), we studied the formation of virus-like particles (VLP) of a gammaretrovirus, the xenotropic murine leukemia virus (MLV)-related virus (XMRV). We show here that, unlike other retroviruses, XMRV CA and CANC do not assemble tubular particles characteristic of mature assembly. The prevention of β-hairpin formation by the deletion of either the N-terminal proline or 10 initial amino acids enabled the assembly of ΔProCANC or Δ10CANC into immature-like spherical particles. Detailed three-dimensional (3D) structural analysis of these particles revealed that below a disordered N-terminal CA layer, the C terminus of CA assembles a typical immature lattice, which is linked by rod-like densities with the RNP.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., IOCB and Gilead Research Center, Prague, Czech Republic.
authors.forename: Romana
authors.initials: R
authors.lastname: Hadravová
authors.forename: Alex
authors.initials: A
authors.lastname: de Marco
authors.forename: Pavel
authors.initials: P
authors.lastname: Ulbrich
authors.forename: Jitka
authors.initials: J
authors.lastname: Stokrová
authors.forename: Michal
authors.initials: M
authors.lastname: Dolezal
authors.forename: Iva
authors.initials: I
authors.lastname: Pichová
authors.forename: Tomás
authors.initials: T
authors.lastname: Ruml
authors.forename: John A G
authors.initials: JA
authors.lastname: Briggs
authors.forename: Michaela
authors.initials: M
authors.lastname: Rumlová
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D020285:Cryoelectron Microscopy; D017931:DNA Primers; D004926:Escherichia coli / Q000648:ultrastructure / Q000821:virology; D005583:Fourier Analysis; D009052:Leukemia Virus, Murine / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D059748:Proteolysis; D017386:Sequence Homology, Amino Acid; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism; D014771:Virion / Q000502:physiology*; D019065:Virus Assembly
chemical_list: D017931:DNA Primers; D014764:Viral Proteins
references.citation: J Virol. 1998 Mar;72(3):2072-8
references.pmid: 9499062
references.citation: J Virol. 2010 Nov;84(22):11729-36
references.pmid: 20810738
references.citation: J Virol. 1995 Oct;69(10):6487-97
references.pmid: 7666550
references.citation: Virus Res. 2011 Jan;155(1):181-8
references.pmid: 20934471
references.citation: EMBO J. 2008 May 7;27(9):1411-20
references.pmid: 18401344
references.citation: Virology. 2003 Mar 30;308(1):83-91
references.pmid: 12706092
references.citation: J Mol Biol. 2009 Sep 11;392(1):100-14
references.pmid: 19527730
references.citation: Prostate. 2008 Feb 15;68(3):306-20
references.pmid: 18163428
references.citation: J Virol. 2003 Mar;77(5):3339-44
references.pmid: 12584360
references.citation: Structure. 2000 Jun 15;8(6):617-28
references.pmid: 10873863
references.citation: PLoS Pathog. 2010 Nov 24;6(11):e1001215
references.pmid: 21151640
references.citation: Science. 1996 Jul 12;273(5272):231-5
references.pmid: 8662505
references.citation: J Virol. 1995 Jun;69(6):3407-19
references.pmid: 7745687
references.citation: J Virol. 1995 Feb;69(2):1093-8
references.pmid: 7815488
references.citation: Proc Natl Acad Sci U S A. 2009 Jul 7;106(27):11090-5
references.pmid: 19549863
references.citation: Virology. 2007 Apr 10;360(2):434-46
references.pmid: 17123565
references.citation: J Struct Biol. 1999 Dec 1;128(1):82-97
references.pmid: 10600563
references.citation: J Virol. 1992 Aug;66(8):4874-83
references.pmid: 1629958
references.citation: Nature. 2000 Sep 21;407(6802):409-13
references.pmid: 11014200
references.citation: EMBO J. 1998 Mar 16;17(6):1555-68
references.pmid: 9501077
references.citation: Nature. 2011 Jan 20;469(7330):424-7
references.pmid: 21248851
references.citation: Virology. 2008 Oct 10;380(1):157-63
references.pmid: 18755489
references.citation: J Virol. 2001 Mar;75(6):2753-64
references.pmid: 11222698
references.citation: Trends Microbiol. 2011 Nov;19(11):525-9
references.pmid: 21978843
references.citation: Proc Natl Acad Sci U S A. 2011 Apr 5;108(14):5771-6
references.pmid: 21436027
references.citation: J Virol. 2001 May;75(9):4407-12
references.pmid: 11287591
references.citation: PLoS One. 2011 Mar 09;6(3):e17592
references.pmid: 21408077
references.citation: J Mol Biol. 2001 Mar 2;306(4):783-97
references.pmid: 11243788
references.citation: J Virol. 1998 Jun;72(6):4798-810
references.pmid: 9573245
references.citation: EMBO J. 2000 Jan 4;19(1):103-13
references.pmid: 10619849
references.citation: J Virol. 2000 Jun;74(12):5395-402
references.pmid: 10823843
references.citation: Retrovirology. 2009 Oct 16;6:92
references.pmid: 19835577
references.citation: Eur J Biochem. 1997 Oct 15;249(2):592-600
references.pmid: 9370371
references.citation: Science. 1999 Jan 1;283(5398):80-3
references.pmid: 9872746
references.citation: PLoS Pathog. 2006 Mar;2(3):e25
references.pmid: 16609730
references.citation: Retrovirology. 2011 Feb 22;8:12
references.pmid: 21342521
references.citation: J Mol Biol. 1999 Aug 13;291(2):491-505
references.pmid: 10438634
references.citation: J Struct Biol. 1996 Jan-Feb;116(1):17-24
references.pmid: 8742718
references.citation: J Mol Biol. 2006 Jan 6;355(1):157-68
references.pmid: 16289202
references.citation: J Virol. 2003 Jun;77(12):7058-66
references.pmid: 12768025
references.citation: J Virol. 2008 Jul;82(14):6788-97
references.pmid: 18448521
references.citation: Science. 2011 Jul 1;333(6038):97-101
references.pmid: 21628392
references.citation: J Virol. 1998 Nov;72(11):9313-7
references.pmid: 9765481
references.citation: J Virol. 2010 Feb;84(4):1977-88
references.pmid: 20007269
references.citation: Curr Opin Struct Biol. 2008 Apr;18(2):203-17
references.pmid: 18406133
references.citation: J Clin Virol. 2008 Nov;43(3):277-83
references.pmid: 18823818
references.citation: J Virol. 2002 Jun;76(11):5452-62
references.pmid: 11991973
references.citation: J Mol Biol. 2008 Mar 7;376(5):1493-508
references.pmid: 18222469
references.citation: J Virol. 2000 Sep;74(18):8452-9
references.pmid: 10954545
references.citation: Retrovirology. 2010 Jul 01;7:57
references.pmid: 20594299
references.citation: Science. 2011 Jul 1;333(6038):94-7
references.pmid: 21628393
references.citation: J Virol. 2001 Jan;75(2):759-71
references.pmid: 11134289
references.citation: J Virol. 2003 Feb;77(3):1772-83
references.pmid: 12525611
references.citation: Retrovirology. 2011 Mar 17;8:20
references.pmid: 21414229
references.citation: Nat Struct Mol Biol. 2004 Jul;11(7):672-5
references.pmid: 15208690
references.citation: Science. 2009 Oct 23;326(5952):585-9
references.pmid: 19815723
references.citation: Virol J. 2010 Sep 13;7:224
references.pmid: 20836869
references.citation: J Virol. 2003 May;77(9):5439-50
references.pmid: 12692245
references.citation: Prostate. 2011 Mar 1;71(4):415-20
references.pmid: 20878949
references.citation: J Virol. 2004 Jan;78(1):52-60
references.pmid: 14671087
references.citation: Nature. 2004 Sep 23;431(7007):481-5
references.pmid: 15386017
references.citation: EMBO J. 2007 Apr 18;26(8):2218-26
references.pmid: 17396149
references.citation: J Struct Biol. 1996 Jan-Feb;116(1):71-6
references.pmid: 8742726
references.citation: Ann Neurol. 2011 Apr;69(4):735-8
references.pmid: 21472770
references.citation: J Virol. 2006 Jul;80(14):7089-99
references.pmid: 16809314
references.citation: Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7299-304
references.pmid: 9636143
references.citation: J Virol. 2011 Dec;85(23):12733-41
references.pmid: 21917964
references.citation: J Virol. 2011 Jul;85(14):7195-202
references.pmid: 21543496