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J Virol. 1987 Feb; 61(2): 454–464.
PMCID: PMC253969

Continuing coevolution of virus and defective interfering particles and of viral genome sequences during undiluted passages: virus mutants exhibiting nearly complete resistance to formerly dominant defective interfering particles.


We quantitatively analyzed the interference interactions between defective interfering (DI) particles and mutants of cloned vesicular stomatitis virus passaged undiluted hundreds of times in BHK-21 cells. DI particles which predominated at different times in these serial passages always interfered most strongly (and very efficiently) with virus isolated a number of passages before the isolation of the DI particles. Virus isolated at the same passage level as the predominant DI particles usually exhibited severalfold resistance to these DI particles. Virus mutants (Sdi- mutants) isolated during subsequent passages always showed increasing resistance to these DI particles, followed by decreasing resistance as new DI particles arose to predominate and exert their own selective pressures on the virus mutant population. It appears that such coevolution of virus and DI particle populations proceeds indefinitely through multiple cycles of selection of virus mutants resistant to a certain DI particle (or DI particle class), followed by mutants resistant to a newly predominant DI particle, etc. At the peak of resistance, virus mutants were isolated which were essentially completely resistant to a particular DI particle; i.e., they were several hundred thousand-fold resistant, and they formed plaques of normal size and numbers in the presence of extremely high multiplicities of the DI particle. However, they were sensitive to interference by other DI particles. Recurring population interactions of this kind can promote rapid virus evolution. Complete sequencing of the N (nucleocapsid) and NS (polymerase associated) genes of numerous Sdi- mutants collected at passage intervals showed very few changes in the NS protein, but the N gene gradually accumulated a series of stable nucleotide and amino acid substitutions, some of which correlated with extensive changes in the Sdi- phenotype. Likewise, the 5' termini (and their complementary plus-strand 3' termini) continued to accumulate extensive base substitutions which were strikingly confined to the first 47 nucleotides. We also observed addition and deletion mutations in noncoding regions of the viral genome at a level suggesting that they probably occur at a high frequency throughout the genome, but usually with lethal or debilitating consequences when they occur in coding regions.

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  • Arnheiter H, Davis NL, Wertz G, Schubert M, Lazzarini RA. Role of the nucleocapsid protein in regulating vesicular stomatitis virus RNA synthesis. Cell. 1985 May;41(1):259–267. [PubMed]
  • Barrett AD, Dimmock NJ. Modulation of a systemic Semliki Forest virus infection in mice by defective interfering virus. J Gen Virol. 1984 Oct;65(Pt 10):1827–1831. [PubMed]
  • Bean WJ, Kawaoka Y, Wood JM, Pearson JE, Webster RG. Characterization of virulent and avirulent A/chicken/Pennsylvania/83 influenza A viruses: potential role of defective interfering RNAs in nature. J Virol. 1985 Apr;54(1):151–160. [PMC free article] [PubMed]
  • Isle HD, Emerson SU. Use of a hybrid infectivity assay to analyze primary transcription of temperature-sensitive mutants of the New Jersey serotype of vesicular stomatitis virus. J Virol. 1982 Jul;43(1):37–40. [PMC free article] [PubMed]
  • Blumberg BM, Giorgi C, Kolakofsky D. N protein of vesicular stomatitis virus selectively encapsidates leader RNA in vitro. Cell. 1983 Feb;32(2):559–567. [PubMed]
  • Blumberg BM, Leppert M, Kolakofsky D. Interaction of VSV leader RNA and nucleocapsid protein may control VSV genome replication. Cell. 1981 Mar;23(3):837–845. [PubMed]
  • Brinton MA, Fernandez AV. A replication-efficient mutant of West Nile virus is insensitive to DI particle interference. Virology. 1983 Aug;129(1):107–115. [PubMed]
  • Cave DR, Hagen FS, Palma EL, Huang AS. Detection of vesicular stomatitis virus RNA and its defective-interfering particles in individual mouse brains. J Virol. 1984 Apr;50(1):86–91. [PMC free article] [PubMed]
  • Cave DR, Hendrickson FM, Huang AS. Defective interfering virus particles modulate virulence. J Virol. 1985 Aug;55(2):366–373. [PMC free article] [PubMed]
  • Clinton GM, Burge BW, Huang AS. Phosphoproteins of vesicular stomatitis virus: identity and interconversion of phosphorylated forms. Virology. 1979 Nov;99(1):84–94. [PubMed]
  • Doyle M, Holland JJ. Prophylaxis and immunization in mice by use of virus-free defective T particles to protect against intracerebral infection by vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2105–2108. [PMC free article] [PubMed]
  • Emerson SU, Wagner RR. Dissociation and reconstitution of the transcriptase and template activities of vesicular stomatitis B and T virions. J Virol. 1972 Aug;10(2):297–309. [PMC free article] [PubMed]
  • Enea V, Zinder ND. Interference resistant mutants of phage f1. Virology. 1982 Oct 15;122(1):222–226. [PubMed]
  • Gallione CJ, Greene JR, Iverson LE, Rose JK. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus N and NS proteins. J Virol. 1981 Aug;39(2):529–535. [PMC free article] [PubMed]
  • Henle W, Henle G. INTERFERENCE OF INACTIVE VIRUS WITH THE PROPAGATION OF VIRUS OF INFLUENZA. Science. 1943 Jul 23;98(2534):87–89. [PubMed]
  • Hill VM, Summers DF. Synthesis of VSV RNPs in vitro by cellular VSV RNPs added to uninfected HeLa cell extracts: VSV protein requirements for replication in vitro. Virology. 1982 Dec;123(2):407–419. [PubMed]
  • Horodyski FM, Holland JJ. Viruses isolated from cells persistently infected with vesicular stomatitis virus show altered interactions with defective interfering particles. J Virol. 1980 Nov;36(2):627–631. [PMC free article] [PubMed]
  • Horodyski FM, Nichol ST, Spindler KR, Holland JJ. Properties of DI particle resistant mutants of vesicular stomatitis virus isolated from persistent infections and from undiluted passages. Cell. 1983 Jul;33(3):801–810. [PubMed]
  • Hsu CH, Morgan EM, Kingsbury DW. Site-specific phosphorylation regulates the transcriptive activity of vesicular stomatitis virus NS protein. J Virol. 1982 Jul;43(1):104–112. [PMC free article] [PubMed]
  • Huang AS, Manders EK. Ribonucleic acid synthesis of vesicular stomatitis virus. IV. Transcription by standard virus in the presence of defective interfering particles. J Virol. 1972 Jun;9(6):909–916. [PMC free article] [PubMed]
  • Isaac CL, Keene JD. RNA polymerase-associated interactions near template promoter sequences of defective interfering particles of vesicular stomatitis virus. J Virol. 1982 Jul;43(1):241–249. [PMC free article] [PubMed]
  • Ito H, Ike Y, Ikuta S, Itakura K. Solid phase synthesis of polynucleotides. VI. Further studies on polystyrene copolymers for the solid support. Nucleic Acids Res. 1982 Mar 11;10(5):1755–1769. [PMC free article] [PubMed]
  • Jacobson S, Pfau CJ. Viral pathogenesis and resistance to defective interfering particles. Nature. 1980 Jan 17;283(5744):311–313. [PubMed]
  • Kawai A, Matsumoto S. Interfering and noninterfering defective particles generated by a rabies small plaque variant virus. Virology. 1977 Jan;76(1):60–71. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lazzarini RA, Keene JD, Schubert M. The origins of defective interfering particles of the negative-strand RNA viruses. Cell. 1981 Oct;26(2 Pt 2):145–154. [PubMed]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [PubMed]
  • MIMS CA. Rift Valley Fever virus in mice. IV. Incomplete virus; its production and properties. Br J Exp Pathol. 1956 Apr;37(2):129–143. [PMC free article] [PubMed]
  • Miyoshi K, Arentzen R, Huang T, Itakura K. Solid-phase synthesis of polynucleotides. IV. Usage of polystyrene resins for the synthesis of polydeoxyribonucleotides by the phosphostriester method. Nucleic Acids Res. 1980 Nov 25;8(22):5507–5517. [PMC free article] [PubMed]
  • O'Hara PJ, Horodyski FM, Nichol ST, Holland JJ. Vesicular stomatitis virus mutants resistant to defective-interfering particles accumulate stable 5'-terminal and fewer 3'-terminal mutations in a stepwise manner. J Virol. 1984 Mar;49(3):793–798. [PMC free article] [PubMed]
  • O'Hara PJ, Nichol ST, Horodyski FM, Holland JJ. Vesicular stomatitis virus defective interfering particles can contain extensive genomic sequence rearrangements and base substitutions. Cell. 1984 Apr;36(4):915–924. [PubMed]
  • Ongrádi J, Cunningham C, Szilágyi JF. Temperature sensitivity of the transcriptase of mutants tsB1 and tsF1 of vesicular stomatitis virus New Jersey is a consequence of mutation affecting polypeptide L. J Gen Virol. 1985 Jul;66(Pt 7):1507–1513. [PubMed]
  • Palma EL, Huang A. Cyclic production of vesicular stomatitis virus caused by defective interfering particles. J Infect Dis. 1974 Apr;129(4):402–410. [PubMed]
  • Palma EL, Perlman SM, Huang AS. Ribonucleic acid synthesis of vesicular stomatitis virus. VI. Correlation of defective particle RNA synthesis with standard RNA replication. J Mol Biol. 1974 May 5;85(1):127–136. [PubMed]
  • Patton JT, Davis NL, Wertz GW. Cell-free synthesis and assembly of vesicular stomatitis virus nucleocapsids. J Virol. 1983 Jan;45(1):155–164. [PMC free article] [PubMed]
  • Patton JT, Davis NL, Wertz GW. N protein alone satisfies the requirement for protein synthesis during RNA replication of vesicular stomatitis virus. J Virol. 1984 Feb;49(2):303–309. [PMC free article] [PubMed]
  • Peluso RW, Moyer SA. Initiation and replication of vesicular stomatitis virus genome RNA in a cell-free system. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3198–3202. [PMC free article] [PubMed]
  • Perrault J. Origin and replication of defective interfering particles. Curr Top Microbiol Immunol. 1981;93:151–207. [PubMed]
  • Perrault J, Clinton GM, McClure MA. RNP template of vesicular stomatitis virus regulates transcription and replication functions. Cell. 1983 Nov;35(1):175–185. [PubMed]
  • Perrault J, Holland JJ. Absence of transcriptase activity or transcription-inhibiting ability in defective interfering particles of vesicular stomatitis virus. Virology. 1972 Oct;50(1):150–170. [PubMed]
  • Popescu M, Lehmann-Grube F. Defective interfering particles in mice infected with lymphocytic choriomeningitis virus. Virology. 1977 Mar;77(1):78–83. [PubMed]
  • Pringle CR. Genetic characteristics of conditional lethal mutants of vesicular stomatitis virus induced by 5-fluorouracil, 5-azacytidine, and ethyl methane sulfonate. J Virol. 1970 May;5(5):559–567. [PMC free article] [PubMed]
  • Reichmann ME, Schnitzlein WM. Defective interfering particles of rhabdoviruses. Curr Top Microbiol Immunol. 1979;86:123–168. [PubMed]
  • Rose JK. Complete intergenic and flanking gene sequences from the genome of vesicular stomatitis virus. Cell. 1980 Feb;19(2):415–421. [PubMed]
  • Schubert M, Harmison GG, Meier E. Primary structure of the vesicular stomatitis virus polymerase (L) gene: evidence for a high frequency of mutations. J Virol. 1984 Aug;51(2):505–514. [PMC free article] [PubMed]
  • Semler BL, Holland JJ. Persistent vesicular stomatitis virus infection mediates base substitutions in viral RNA termini. J Virol. 1979 Nov;32(2):420–428. [PMC free article] [PubMed]
  • Spindler KR, Horodyski FM, Holland JJ. High multiplicities of infection favor rapid and random evolution of vesicular stomatitis virus. Virology. 1982 May;119(1):96–108. [PubMed]
  • Stamminger G, Lazzarini RA. Analysis of the RNA of defective VSV particles. Cell. 1974 Sep;3(1):85–93. [PubMed]
  • Testa D, Banerjee AK. Initiation of RNA synthesis in vitro by vesicular stomatitis virus. Role of ATP. J Biol Chem. 1979 Mar 25;254(6):2053–2058. [PubMed]
  • VON MAGNUS P. Incomplete forms of influenza virus. Adv Virus Res. 1954;2:59–79. [PubMed]
  • Weiss B, Schlesinger S. Defective interfering particles of Sindbis virus do not interfere with the homologous virus obtained from persistently infected BHK cells but do interfere with Semliki Forest virus. J Virol. 1981 Feb;37(2):840–844. [PMC free article] [PubMed]
  • Wilusz J, Youngner JS, Keene JD. Base mutations in the terminal noncoding regions of the genome of vesicular stomatitis virus isolated from persistent infections of L cells. Virology. 1985 Jan 30;140(2):249–256. [PubMed]

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