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PMID: 10864650 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Poliovirus requires a precise 5' end for efficient positive-strand RNA synthesis.

Journal of virology ·Vol. 74 ·No. 14 ·2000-07-00 ·Pages 6394-400

Herold J, Andino R

Abstract

Poliovirus infectious RNA can be synthesized in vitro using phage DNA-dependent RNA-polymerases. These synthetic transcripts contain several extra nucleotides at the 5' end, which are deleted during replication to generate authentic viral genomes. We removed those 5'-end extra nucleotides utilizing a hammerhead ribozyme to produce transcripts with accurate 5' ends. These transcripts replicate substantially more rapidly in cell culture, demonstrating no lag before replication; they also replicate more efficiently in Xenopus laevis oocytes and in in vitro translation-replication cell extracts. In both systems, an exact 5' end is necessary for synthesis of positive-strand RNA but not negative-strand RNA.

MeSH Terms
Animals Cell Extracts Cell Line Humans In Vitro Techniques Oocytes/virology Poliovirus/genetics,metabolism,physiology RNA, Catalytic/metabolism RNA, Viral/biosynthesis,chemistry Virus Replication Xenopus laevis
Chemicals
Cell Extracts RNA, Catalytic RNA, Viral
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Herold J
Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, California 94143-0414, USA.
Andino R
References (36)
36 references, click to expand
  1. Effects of mutations in poliovirus 3Dpol on RNA polymerase activity and on polyprotein cleavage.
    J Virol. 1989 Nov;63(11):4866-74 PMID: 2552159
  2. Infectious transcripts and cDNA clones of RNA viruses.
    Virology. 1994 Feb;198(2):415-26 PMID: 8291226
  3. Proteolytic processing of polyproteins in the replication of RNA viruses.
    Biochemistry. 1989 Dec 26;28(26):9881-90 PMID: 2695162
  4. The 5'-terminal nucleotides of hepatitis A virus RNA, but not poliovirus RNA, are required for infectivity.
    J Virol. 1991 May;65(5):2757-60 PMID: 1850050
  5. Cell-free, de novo synthesis of poliovirus.
    Science. 1991 Dec 13;254(5038):1647-51 PMID: 1661029
  6. Rescue of defective poliovirus RNA replication by 3AB-containing precursor polyproteins.
    J Virol. 1998 Sep;72(9):7191-200 PMID: 9696813
  7. Genetic analysis of picornaviruses.
    Curr Opin Genet Dev. 1992 Feb;2(1):64-70 PMID: 1321679
  8. Assays for poliovirus polymerase, 3D(Pol), and authentic RNA replication in HeLa S10 extracts.
    Methods Enzymol. 1996;275:35-57 PMID: 9026649
  9. A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA.
    Cell. 1990 Oct 19;63(2):369-80 PMID: 2170027
  10. Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3-7c inhibit cleavage at glutamine-glycine pairs.
    Proc Natl Acad Sci U S A. 1982 Jul;79(13):3973-7 PMID: 6287457
  11. Cloned poliovirus complementary DNA is infectious in mammalian cells.
    Science. 1981 Nov 20;214(4523):916-9 PMID: 6272391
  12. Isolation of poliovirus 2C mutants defective in viral RNA synthesis.
    J Virol. 1988 Nov;62(11):4016-21 PMID: 2845120
  13. Improved method for detecting poliovirus negative strands used to demonstrate specificity of positive-strand encapsidation and the ratio of positive to negative strands in infected cells.
    J Virol. 1991 Jun;65(6):3384-7 PMID: 1851886
  14. Complete nucleotide sequence of infectious Coxsackievirus B3 cDNA: two initial 5' uridine residues are regained during plus-strand RNA synthesis.
    J Virol. 1990 Apr;64(4):1573-83 PMID: 2157045
  15. Poliovirus RNA synthesis utilizes an RNP complex formed around the 5'-end of viral RNA.
    EMBO J. 1993 Sep;12(9):3587-98 PMID: 8253083
  16. Hybridization of nucleic acids immobilized on solid supports.
    Anal Biochem. 1984 May 1;138(2):267-84 PMID: 6204550
  17. Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase.
    Proc Natl Acad Sci U S A. 1986 Apr;83(8):2330-4 PMID: 3010307
  18. Switch from translation to RNA replication in a positive-stranded RNA virus.
    Genes Dev. 1998 Aug 1;12(15):2293-304 PMID: 9694795
  19. The structure, function and application of the hammerhead ribozyme.
    Eur J Biochem. 1997 Apr 1;245(1):1-16 PMID: 9128718
  20. Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases.
    Nucleic Acids Res. 1988 Oct 25;16(20):9677-86 PMID: 2460825
  21. Role of a viral membrane polypeptide in strand-specific initiation of poliovirus RNA synthesis.
    J Virol. 1991 May;65(5):2647-54 PMID: 1850038
  22. Cloning and synthesis of infectious cardiovirus RNAs containing short, discrete poly(C) tracts.
    J Virol. 1989 Apr;63(4):1822-6 PMID: 2538661
  23. Genetic complementation among poliovirus mutants derived from an infectious cDNA clone.
    J Virol. 1986 Dec;60(3):1040-9 PMID: 3023656
  24. Multicomponent RNA plant virus infection derived from cloned viral cDNA.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):7066-70 PMID: 16593527
  25. Replication of poliovirus in Xenopus oocytes requires two human factors.
    EMBO J. 1996 Nov 1;15(21):5988-98 PMID: 8918476
  26. Protein-primed RNA synthesis by purified poliovirus RNA polymerase.
    Nature. 1998 May 21;393(6682):280-4 PMID: 9607767
  27. Mutational analysis of the genome-linked protein VPg of poliovirus.
    J Virol. 1988 Nov;62(11):4207-15 PMID: 2845132
  28. Translating ribosomes inhibit poliovirus negative-strand RNA synthesis.
    J Virol. 1999 Dec;73(12):10104-12 PMID: 10559325
  29. Role of 3'-end sequences in infectivity of poliovirus transcripts made in vitro.
    J Virol. 1989 Jan;63(1):467-70 PMID: 2535751
  30. A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein.
    Cell. 1986 Jun 6;45(5):761-70 PMID: 3011278
  31. Effective amplification of 20-kb DNA by reverse transcription PCR.
    Anal Biochem. 1997 Oct 1;252(1):62-70 PMID: 9324942
  32. Interaction of poliovirus polypeptide 3CDpro with the 5' and 3' termini of the poliovirus genome. Identification of viral and cellular cofactors needed for efficient binding.
    J Biol Chem. 1994 Oct 28;269(43):27004-14 PMID: 7929441
  33. Crystallographic analyses of chemically synthesized modified hammerhead RNA sequences as a general approach toward understanding ribozyme structure and function.
    Methods Mol Biol. 1997;74:387-91 PMID: 9204454
  34. Complete replication of poliovirus in vitro: preinitiation RNA replication complexes require soluble cellular factors for the synthesis of VPg-linked RNA.
    J Virol. 1995 Sep;69(9):5516-27 PMID: 7636997
  35. Translation and replication properties of the human rhinovirus genome in vivo and in vitro.
    Virology. 1997 Mar 3;229(1):90-7 PMID: 9123881
  36. The structure of poliovirus replicative form.
    Nucleic Acids Res. 1980 Mar 25;8(6):1217-29 PMID: 6253931
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.14.6394-6400.2000
Published
2000-07-00
Pages
6394-400
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC112146
Subset
IM
Grants
NIAID NIH HHS · N01AI40085 · United States
External Links
PubMed source DOI: 10.1128/jvi.74.14.6394-6400.2000 PMC Full Text View journal details
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