title: Biological differences between vesicular stomatitis virus Indiana and New Jersey serotype glycoproteins: identification of amino acid residues modulating pH-dependent infectivity.
abstract: We previously generated recombinant vesicular stomatitis viruses (VSV) based on the Indiana serotype genome which contained either the homologous glycoprotein gene from the Indiana serotype (VSIV-GI) or the heterologous glycoprotein gene from the New Jersey serotype (VSIV-GNJ). The virus expressing the GNJ gene was more pathogenic than the parental VSIV-GI virus in swine, a natural host (26). For the present study, we investigated the biological differences between the GI and GNJ proteins that may be related to the differences in pathogenesis between VSIV-GI and VSIV-GNJ. We show that the capacities of viruses with either the GNJ or GI glycoprotein to infect cultured cells differ depending on the pH. VSIV-GNJ could infect cells at acidic pHs, while the infectivity of VSIV-GI was severely reduced. VSIV-GNJ infection was also more sensitive to inhibition by ammonium chloride, indicating that the GNJ protein had a lower pH threshold for membrane fusion. We applied selective pressure to VSIV-GI by growing it at successively lower pH values and isolated variant viruses in which we identified amino acid changes that conferred low-pH-resistant infectivity. Repeated passage in cell culture at pH 6.8 resulted in the selection of a VSIV-GI variant (VSIV-6.8) that was similar to VSIV-GNJ regarding its pH- and ammonium chloride-dependent infectivity. Sequence analysis of VSIV-6.8 revealed that it had a single amino acid substitution in the amino-terminal region of the glycoprotein (F18L). This alteration was shown to be responsible for the observed phenotype by site-directed mutagenesis of a VSIV-GI full-length cDNA and analysis of the recovered engineered virus. A further adaptation of VSIV-6.8 to pHs 6.6 and 6.4 resulted in additional amino acid substitutions in areas of the glycoprotein that were not previously implicated in attachment or fusion.
authors.affiliation: Department of Microbiology, University of Alabama School of Medicine, BBRB Box 17, Room 366, 845 19th St. South, Birmingham, AL 35294, USA.
authors.forename: Isidoro
authors.initials: I
authors.lastname: Martinez
authors.forename: Gail W
authors.initials: GW
authors.lastname: Wertz
mesh_terms: D019943:Amino Acid Substitution; D000596:Amino Acids / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004252:DNA Mutational Analysis; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D016297:Mutagenesis, Site-Directed; D012641:Selection, Genetic; D012703:Serotyping; D014709:Vero Cells; D014721:Vesicular stomatitis Indiana virus / Q000145:classification / Q000235:genetics* / Q000502:physiology*; D018116:Vesiculovirus; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D010948:Viral Plaque Assay
chemical_list: D000596:Amino Acids; D006023:Glycoproteins; D014759:Viral Envelope Proteins
references.citation: J Virol. 2002 Apr;76(8):3756-64
references.pmid: 11907215
references.citation: Virology. 1989 Jun;170(2):392-407
references.pmid: 2471352
references.citation: J Gen Virol. 1972 Sep;16(3):391-8
references.pmid: 4342823
references.citation: J Virol. 1981 Aug;39(2):519-28
references.pmid: 6268840
references.citation: J Virol. 2003 Dec;77(23):12807-18
references.pmid: 14610202
references.citation: J Virol. 1972 Dec;10(6):1231-5
references.pmid: 4118869
references.citation: J Gen Virol. 1987 Sep;68 ( Pt 9):2359-69
references.pmid: 2821175
references.citation: Virology. 1998 Mar 1;242(1):39-50
references.pmid: 9501039
references.citation: Biochemistry. 1996 Apr 2;35(13):4084-93
references.pmid: 8672443
references.citation: Cell. 1983 Feb;32(2):639-46
references.pmid: 6297804
references.citation: Am J Vet Res. 2001 Apr;62(4):516-20
references.pmid: 11327457
references.citation: J Biol Chem. 1995 Jul 21;270(29):17575-81
references.pmid: 7615563
references.citation: Virology. 1996 Mar 1;217(1):49-57
references.pmid: 8599235
references.citation: J Mol Biol. 1982 Apr 15;156(3):609-31
references.pmid: 6288961
references.citation: Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8388-92
references.pmid: 7667300
references.citation: J Cell Biol. 1981 Jun;89(3):674-9
references.pmid: 6265470
references.citation: J Biol Chem. 2003 Apr 18;278(16):13789-94
references.pmid: 12571240
references.citation: J Virol. 2002 Dec;76(23):12300-11
references.pmid: 12414970
references.citation: J Cell Biol. 1987 Nov;105(5):1957-69
references.pmid: 2824524
references.citation: Biochemistry. 1990 Feb 6;29(5):1303-8
references.pmid: 2157487
references.citation: J Virol. 1995 Mar;69(3):1435-43
references.pmid: 7853475
references.citation: Virus Res. 2002 May 10;85(2):211-9
references.pmid: 12034487
references.citation: J Virol. 1983 Apr;46(1):162-9
references.pmid: 6298453
references.citation: Biochemistry. 1983 Dec 6;22(25):5790-6
references.pmid: 6661413
references.citation: Biochemistry. 1997 Jul 22;36(29):8890-6
references.pmid: 9220976
references.citation: J Virol. 1993 Jul;67(7):4070-7
references.pmid: 8389917
references.citation: Intervirology. 1991;32(3):193-7
references.pmid: 1645706
references.citation: J Virol. 2003 Jul;77(14):8039-47
references.pmid: 12829843
references.citation: J Virol. 2000 Mar;74(5):2239-46
references.pmid: 10666254
references.citation: J Biol Chem. 2001 Jan 5;276(1):62-7
references.pmid: 11024041
references.citation: Vet J. 1999 May;157(3):239-60
references.pmid: 10328837
references.citation: Virology. 1998 Jan 20;240(2):349-58
references.pmid: 9454708
references.citation: J Virol. 1999 Jun;73(6):4705-12
references.pmid: 10233930
references.citation: J Biol Chem. 1990 Apr 25;265(12):6879-83
references.pmid: 2157712
references.citation: J Am Vet Med Assoc. 1999 Nov 1;215(9):1259-62
references.pmid: 10553435
references.citation: Virology. 2000 May 25;271(1):171-81
references.pmid: 10814582
references.citation: J Virol. 2003 Mar;77(6):3785-98
references.pmid: 12610153
references.citation: J Biol Chem. 1988 Apr 5;263(10):4749-53
references.pmid: 2832405
references.citation: J Am Vet Med Assoc. 1997 Sep 1;211(5):556-60
references.pmid: 9290819
references.citation: Biochemistry. 1992 Oct 20;31(41):10108-13
references.pmid: 1327132
references.citation: J Virol. 1986 Feb;57(2):603-13
references.pmid: 3003392
references.citation: J Virol. 1985 Jan;53(1):319-23
references.pmid: 2981356
references.citation: J Virol. 1986 Aug;59(2):486-9
references.pmid: 3016308
references.citation: Cell. 1985 Feb;40(2):431-9
references.pmid: 3967299
references.citation: J Virol. 1995 Sep;69(9):5528-34
references.pmid: 7543584
references.citation: J Virol. 1994 Apr;68(4):2186-93
references.pmid: 8139003
references.citation: Virology. 1999 Mar 30;256(1):119-29
references.pmid: 10087232