The Efficient Packaging of Venezuelan Equine Encephalitis Virus-specific RNAs into Viral Particles is Determined by NsP1-3 Synthesis

Overview

Journal Virology

Specialty Microbiology

Date 2005 Oct 22

PMID 16239019

Citations 47

Authors

Eugenia Volkova

Rodion Gorchakov

Ilya Frolov

Affiliations

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Abstract

Alphaviruses are regarded as attractive systems for expression of heterologous genes and development of recombinant vaccines. Venezuelan equine encephalitis virus (VEE)-based vectors are particularly promising because of their specificity to lymphoid tissues and strong resistance to interferon. To improve understanding of the VEE genome packaging and optimize application of this virus as a vector, we analyzed in more detail the mechanism of packaging of the VEE-specific RNAs. The presence of the RNAs in the VEE particles during serial passaging in tissue culture was found to depend not only on the presence of packaging signal(s), but also on the ability of these RNAs to express in cis nsP1, nsP2 and nsP3 in the form of a P123 precursor. Packaging of VEE genomes into infectious virions was also found to be more efficient compared to that of Sindbis virus, in spite of lower levels of RNA replication and structural protein production.

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References

Perri S, Greer C, Thudium K, Doe B, Legg H, Liu H . An alphavirus replicon particle chimera derived from venezuelan equine encephalitis and sindbis viruses is a potent gene-based vaccine delivery vector. J Virol. 2003; 77(19):10394-403. PMC: 228391. DOI: 10.1128/jvi.77.19.10394-10403.2003. View

Gorchakov R, Frolova E, Williams B, Rice C, Frolov I . PKR-dependent and -independent mechanisms are involved in translational shutoff during Sindbis virus infection. J Virol. 2004; 78(16):8455-67. PMC: 479073. DOI: 10.1128/JVI.78.16.8455-8467.2004. View

Weaver S, Barrett A . Transmission cycles, host range, evolution and emergence of arboviral disease. Nat Rev Microbiol. 2004; 2(10):789-801. PMC: 7097645. DOI: 10.1038/nrmicro1006. View

Sarkar S, Sen G . Novel functions of proteins encoded by viral stress-inducible genes. Pharmacol Ther. 2004; 103(3):245-59. DOI: 10.1016/j.pharmthera.2004.07.007. View

Rice C, Strauss J . Nucleotide sequence of the 26S mRNA of Sindbis virus and deduced sequence of the encoded virus structural proteins. Proc Natl Acad Sci U S A. 1981; 78(4):2062-6. PMC: 319283. DOI: 10.1073/pnas.78.4.2062. View

Lehtovaara P, Soderlund H, Keranen S, Pettersson R, Kaariainen L . 18S defective interfering RNA of Semliki Forest virus contains a triplicated linear repeat. Proc Natl Acad Sci U S A. 1981; 78(9):5353-7. PMC: 348743. DOI: 10.1073/pnas.78.9.5353. View

Monroe S, Ou J, Rice C, Schlesinger S, Strauss E, Strauss J . Sequence analysis of cDNA's derived from the RNA of Sindbis virions and of defective interfering particles. J Virol. 1982; 41(1):153-62. PMC: 256736. DOI: 10.1128/JVI.41.1.153-162.1982. View

Lehtovaara P, Soderlund H, Keranen S, Pettersson R, Kaariainen L . Extreme ends of the genome are conserved and rearranged in the defective interfering RNAs of Semliki Forest virus. J Mol Biol. 1982; 156(4):731-48. DOI: 10.1016/0022-2836(82)90139-5. View

Monroe S, Schlesinger S . RNAs from two independently isolated defective interfering particles of Sindbis virus contain a cellular tRNA sequence at their 5' ends. Proc Natl Acad Sci U S A. 1983; 80(11):3279-83. PMC: 394024. DOI: 10.1073/pnas.80.11.3279. View

10.

Monroe S, Schlesinger S . Common and distinct regions of defective-interfering RNAs of Sindbis virus. J Virol. 1984; 49(3):865-72. PMC: 255548. DOI: 10.1128/JVI.49.3.865-872.1984. View

11.

Strauss E, Rice C, Strauss J . Complete nucleotide sequence of the genomic RNA of Sindbis virus. Virology. 1984; 133(1):92-110. DOI: 10.1016/0042-6822(84)90428-8. View

12.

Tsiang M, Monroe S, Schlesinger S . Studies of defective interfering RNAs of Sindbis virus with and without tRNAAsp sequences at their 5' termini. J Virol. 1985; 54(1):38-44. PMC: 254757. DOI: 10.1128/JVI.54.1.38-44.1985. View

13.

Kinney R, Johnson B, Welch J, Tsuchiya K, TRENT D . The full-length nucleotide sequences of the virulent Trinidad donkey strain of Venezuelan equine encephalitis virus and its attenuated vaccine derivative, strain TC-83. Virology. 1989; 170(1):19-30. DOI: 10.1016/0042-6822(89)90347-4. View

14.

Weiss B, Nitschko H, Ghattas I, Wright R, Schlesinger S . Evidence for specificity in the encapsidation of Sindbis virus RNAs. J Virol. 1989; 63(12):5310-8. PMC: 251197. DOI: 10.1128/JVI.63.12.5310-5318.1989. View

15.

Lemm J, Durbin R, Stollar V, Rice C . Mutations which alter the level or structure of nsP4 can affect the efficiency of Sindbis virus replication in a host-dependent manner. J Virol. 1990; 64(6):3001-11. PMC: 249484. DOI: 10.1128/JVI.64.6.3001-3011.1990. View

16.

Weiss B, Wright R, Schlesinger S . Complementation between Sindbis viral RNAs produces infectious particles with a bipartite genome. Proc Natl Acad Sci U S A. 1991; 88(8):3253-7. PMC: 51424. DOI: 10.1073/pnas.88.8.3253. View

17.

Liljestrom P, Lusa S, Huylebroeck D, Garoff H . In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. J Virol. 1991; 65(8):4107-13. PMC: 248843. DOI: 10.1128/JVI.65.8.4107-4113.1991. View

18.

Liljestrom P, Garoff H . A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology (N Y). 1991; 9(12):1356-61. DOI: 10.1038/nbt1291-1356. View

19.

Lemm J, Rice C . Roles of nonstructural polyproteins and cleavage products in regulating Sindbis virus RNA replication and transcription. J Virol. 1993; 67(4):1916-26. PMC: 240259. DOI: 10.1128/JVI.67.4.1916-1926.1993. View

20.

Bredenbeek P, Frolov I, Rice C, Schlesinger S . Sindbis virus expression vectors: packaging of RNA replicons by using defective helper RNAs. J Virol. 1993; 67(11):6439-46. PMC: 238079. DOI: 10.1128/JVI.67.11.6439-6446.1993. View