Detection of Virus-specific RNA-dependent RNA Polymerase Activity in Extracts from Cells Infected with Lymphocytic Choriomeningitis Virus: in Vitro Synthesis of Full-length Viral RNA Species

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1989 May 1

PMID 2704071

Citations 21

Authors

F V Fuller-Pace

P J Southern

Affiliations

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Abstract

We have developed an in vitro assay for the lymphocytic choriomeningitis virus (LCMV) RNA-dependent RNA polymerase with ribonucleoprotein complexes extracted from acutely infected tissue culture cells. The RNA products synthesized in vitro corresponded in size to the full-length genomic L and S RNAs and subgenomic NP and GP mRNAs normally produced in vivo during acute LCMV infection. In a temporal analysis spanning the first 72 h of acute infection, the in vitro polymerase activity of ribonucleoprotein complexes was maximal at 16 h and declined significantly at later times. In contrast, the intracellular levels of the viral L protein (the putative polymerase protein) appeared to be maximal at 48 to 72 h postinfection. Our results suggest that the accumulation of L protein correlates with reduced viral replication and transcription at later times in acute infection and may be involved in the transition from acute to persistent LCMV infection.

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References

Riviere Y, Ahmed R, Southern P, Buchmeier M, Oldstone M . Genetic mapping of lymphocytic choriomeningitis virus pathogenicity: virulence in guinea pigs is associated with the L RNA segment. J Virol. 1985; 55(3):704-9. PMC: 255050. DOI: 10.1128/JVI.55.3.704-709.1985. View

Rigby P, Dieckmann M, Rhodes C, Berg P . Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977; 113(1):237-51. DOI: 10.1016/0022-2836(77)90052-3. View

Bruns M, Zeller W, Rohdewohld H, Lehmann-Grube F . Lymphocytic choriomeningitis virus. IX. Properties of the nucleocapsid. Virology. 1986; 151(1):77-85. DOI: 10.1016/0042-6822(86)90105-4. View

Moyer S, Baker S, Lessard J . Tubulin: a factor necessary for the synthesis of both Sendai virus and vesicular stomatitis virus RNAs. Proc Natl Acad Sci U S A. 1986; 83(15):5405-9. PMC: 386295. DOI: 10.1073/pnas.83.15.5405. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

PEDERSEN I . Lymphocytic choriomeningitis virus RNAs. Nat New Biol. 1971; 234(47):112-4. DOI: 10.1038/newbio234112a0. View

McMaster G, Carmichael G . Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977; 74(11):4835-8. PMC: 432050. DOI: 10.1073/pnas.74.11.4835. View

Vezza A, Clewley J, Gard G, Abraham N, Compans R, Bishop D . Virion RNA species of the arenaviruses Pichinde, Tacaribe, and Tamiami. J Virol. 1978; 26(2):485-97. PMC: 354086. DOI: 10.1128/JVI.26.2.485-497.1978. View

Buchmeier M, Elder J, Oldstone M . Protein structure of lymphocytic choriomeningitis virus: identification of the virus structural and cell associated polypeptides. Virology. 1978; 89(1):133-45. DOI: 10.1016/0042-6822(78)90047-8. View

10.

Vogelstein B, Gillespie D . Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979; 76(2):615-9. PMC: 382999. DOI: 10.1073/pnas.76.2.615. View

11.

Leung W, Leung M, Rawls W . Distinctive RNA transcriptase, polyadenylic acid polymerase, and polyuridylic acid polymerase activities associated with Pichinde virus. J Virol. 1979; 30(1):98-107. PMC: 353303. DOI: 10.1128/JVI.30.1.98-107.1979. View

12.

Buchmeier M, Oldstone M . Protein structure of lymphocytic choriomeningitis virus: evidence for a cell-associated precursor of the virion glycopeptides. Virology. 1979; 99(1):111-20. DOI: 10.1016/0042-6822(79)90042-4. View

13.

Thomas P . Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980; 77(9):5201-5. PMC: 350025. DOI: 10.1073/pnas.77.9.5201. View

14.

Buchmeier M, Welsh R, Dutko F, Oldstone M . The virology and immunobiology of lymphocytic choriomeningitis virus infection. Adv Immunol. 1980; 30:275-331. DOI: 10.1016/s0065-2776(08)60197-2. View

15.

Smith G, Summers M . The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Anal Biochem. 1980; 109(1):123-9. DOI: 10.1016/0003-2697(80)90019-6. View

16.

HILL V, Summers D . 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; 123(2):407-19. DOI: 10.1016/0042-6822(82)90273-2. View

17.

Harnish D, Dimock K, Bishop D, Rawls W . Gene mapping in Pichinde virus: assignment of viral polypeptides to genomic L and S RNAs. J Virol. 1983; 46(2):638-41. PMC: 255169. DOI: 10.1128/JVI.46.2.638-641.1983. View

18.

Lehmann-Grube F . Portraits of viruses: arenaviruses. Intervirology. 1984; 22(3):121-45. DOI: 10.1159/000149543. View

19.

Morrow C, Gibbons G, Dasgupta A . The host protein required for in vitro replication of poliovirus is a protein kinase that phosphorylates eukaryotic initiation factor-2. Cell. 1985; 40(4):913-21. DOI: 10.1016/0092-8674(85)90351-4. View

20.

Compans R, Bishop D . Biochemistry of arenaviruses. Curr Top Microbiol Immunol. 1985; 114:153-75. DOI: 10.1007/978-3-642-70227-3_4. View