title: Hypervariable domain of nonstructural protein nsP3 of Venezuelan equine encephalitis virus determines cell-specific mode of virus replication.
abstract: Venezuelan equine encephalitis virus (VEEV) is one of the most pathogenic members of the Alphavirus genus in the Togaviridae family. This genus is divided into the Old World and New World alphaviruses, which demonstrate profound differences in pathogenesis, replication, and virus-host interactions. VEEV is a representative member of the New World alphaviruses. The biology of this virus is still insufficiently understood, particularly the function of its nonstructural proteins in RNA replication and modification of the intracellular environment. One of these nonstructural proteins, nsP3, contains a hypervariable domain (HVD), which demonstrates very low overall similarity between different alphaviruses, suggesting the possibility of its function in virus adaptation to different hosts and vectors. The results of our study demonstrate the following. (i) Phosphorylation of the VEEV nsP3-specific HVD does not play a critical role in virus replication in cells of vertebrate origin but is important for virus replication in mosquito cells. (ii) The VEEV HVD is not required for viral RNA replication in the highly permissive BHK-21 cell line. In fact, it can be either completely deleted or replaced by a heterologous protein sequence. These variants require only one or two additional adaptive mutations in nsP3 and/or nsP2 proteins to achieve an efficiently replicating phenotype. (iii) However, the carboxy-terminal repeat in the VEEV HVD is indispensable for VEEV replication in the cell lines other than BHK-21 and plays a critical role in formation of VEEV-specific cytoplasmic protein complexes. Natural VEEV variants retain at least one of the repeated elements in their nsP3 HVDs.
authors: Department of Microbiology, University of Alabama, Birmingham, Alabama, USA.
authors: Niall J
authors: NJ
authors: Foy
authors: Maryna
authors: M
authors: Akhrymuk
authors: Alexander V
authors: AV
authors: Shustov
authors: Elena I
authors: EI
authors: Frolova
authors: Ilya
authors: I
authors: Frolov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D006224:Cricetinae; D009033:Culicidae; D018274:Electroporation; D004666:Encephalitis Virus, Venezuelan Equine / Q000235:genetics* / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D014644:Genetic Variation; D051379:Mice; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D010766:Phosphorylation; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D013045:Species Specificity; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics* / Q000502:physiology
chemical_list: D000911:Antibodies, Monoclonal; D017361:Viral Nonstructural Proteins; C089383:nsp3 protein, alphavirus
references: Virology. 1989 Jul;171(1):280-4
references: 2525839
references: J Virol. 2006 Jun;80(12):5686-96
references: 16731907
references: FEBS Lett. 1999 Apr 1;448(1):19-22
references: 10217401
references: J Virol. 2012 Oct;86(19):10873-9
references: 22837213
references: Protein Expr Purif. 2011 Oct;79(2):277-84
references: 21693190
references: Virology. 2009 Feb 5;384(1):201-8
references: 19036396
references: J Virol. 1987 Dec;61(12):3809-19
references: 3479621
references: J Virol. 1994 Mar;68(3):1874-85
references: 8107248
references: J Virol. 1993 Apr;67(4):1916-26
references: 8445717
references: J Virol. 2008 Oct;82(20):10088-101
references: 18684830
references: J Virol. 2011 Apr;85(7):3449-60
references: 21248049
references: EMBO J. 1994 Jun 15;13(12):2925-34
references: 7517863
references: Microbiol Rev. 1994 Sep;58(3):491-562
references: 7968923
references: J Biol Chem. 2006 Oct 6;281(40):30269-78
references: 16895903
references: J Mol Graph Model. 2010 Aug 24;29(1):46-53
references: 20483643
references: J Virol. 2007 Jan;81(2):872-83
references: 17093195
references: Virology. 1996 Apr 15;218(2):352-61
references: 8610462
references: Protein Expr Purif. 2009 Mar;64(1):89-97
references: 19013248
references: J Virol. 2002 Nov;76(22):11254-64
references: 12388685
references: J Virol. 2012 Jul;86(13):7180-91
references: 22514352
references: Structure. 2006 Sep;14(9):1449-58
references: 16962975
references: Nat Rev Microbiol. 2004 Oct;2(10):789-801
references: 15378043
references: J Virol. 2005 Aug;79(15):9397-409
references: 16014903
references: Mol Biol Cell. 2012 Dec;23(24):4701-12
references: 23087212
references: J Virol. 2012 Jan;86(1):553-65
references: 22031949
references: J Virol. 1991 Aug;65(8):4107-13
references: 2072446
references: J Virol. 2010 Oct;84(19):10004-15
references: 20668087
references: J Biol Chem. 2000 Dec 1;275(48):37853-9
references: 10984480
references: J Gen Virol. 2007 Jul;88(Pt 7):1967-1976
references: 17554030
references: J Biol Chem. 2000 Sep 8;275(36):27775-83
references: 10851234
references: J Gen Virol. 2006 Apr;87(Pt 4):937-947
references: 16528043
references: J Virol. 2009 Jul;83(13):6534-45
references: 19386706
references: J Gen Virol. 1991 Jan;72 ( Pt 1):195-9
references: 1824948
references: J Biol Chem. 2001 Feb 23;276(8):5745-52
references: 11104756
references: J Biol Chem. 2001 Aug 17;276(33):30786-93
references: 11410598
references: Virology. 1990 Jul;177(1):54-64
references: 2141206
references: J Virol. 1997 Jan;71(1):392-7
references: 8985362
references: Virology. 1990 Jan;174(1):43-52
references: 2136781
references: PLoS Pathog. 2011 Nov;7(11):e1002383
references: 22114558
references: J Virol. 1999 May;73(5):3854-65
references: 10196280
references: Euro Surveill. 2007 Nov 22;12(11):E071122.2
references: 18053561
references: J Virol. 2007 Apr;81(8):4371-3
references: 17287268
references: Future Microbiol. 2009 Sep;4(7):837-56
references: 19722838
references: J Gen Virol. 2003 Jun;84(Pt 6):1525-1533
references: 12771422
references: J Virol. 1994 Sep;68(9):5781-91
references: 8057460
references: J Virol. 1990 Jun;64(6):3001-11
references: 2159558
references: Virology. 2011 Feb 20;410(2):353-9
references: 21185049
references: Virology. 1989 Jul;171(1):189-204
references: 2525837
references: Proc Natl Acad Sci U S A. 2004 Jun 22;101(25):9429-34
references: 15197279
references: J Virol. 2006 Apr;80(8):4122-34
references: 16571828
references: J Virol. 2003 Dec;77(24):13106-16
references: 14645567
references: J Virol. 2007 Mar;81(5):2472-84
references: 17108023
references: J Virol. 2006 Jan;80(1):360-71
references: 16352561
references: PLoS One. 2011;6(7):e22336
references: 21811589
references: J Virol. 2013 Feb;87(4):2023-35
references: 23221545
references: J Virol. 2013 Feb;87(4):1997-2010
references: 23221551
references: J Mol Graph Model. 2010 Nov;29(3):347-53
references: 21036084
references: J Virol. 2013 Apr;87(8):4202-13
references: 23365438
references: Virology. 1994 Jul;202(1):224-32
references: 7912020
references: Virology. 2009 Jun 5;388(2):305-14
references: 19395054
references: Virology. 1984 Feb;133(1):92-110
references: 6322438
references: J Biol Chem. 2000 Jun 9;275(23):17281-7
references: 10748213
references: N Engl J Med. 2007 Feb 22;356(8):769-71
references: 17314335
references: J Virol. 2010 Nov;84(22):11679-95
references: 20826696
references: Euro Surveill. 2007 Sep 06;12(9):E070906.1
references: 17900424
references: J Virol. 2010 Jul;84(13):6720-32
references: 20392851
references: Proc Natl Acad Sci U S A. 2012 Oct 9;109(41):16534-9
references: 23010928
references: J Virol. 1994 Sep;68(9):5804-10
references: 8057461