Home LiteratureArticle Details
PMID: 3018300 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Initiation of poliovirus plus-strand RNA synthesis in a membrane complex of infected HeLa cells.

Journal of virology ·Vol. 60 ·No. 1 ·1986-10-00 ·Pages 43-53

Takeda N, Kuhn RJ, Yang CF, Takegami T, Wimmer E

Abstract

An in vitro poliovirus RNA-synthesizing system derived from a crude membrane fraction of infected HeLa cells was used to analyze the mechanism of initiation of poliovirus plus-strand RNA synthesis. This system contains an activity that synthesizes the nucleotidyl proteins VPg-pU and VPg-pUpU. These molecules represent the 5'-terminal structure of nascent RNA molecules and of virion RNA. The membranous replication complex is also capable of synthesizing nucleotidyl proteins containing nine or more of the poliovirus 5'-proximal nucleotides as assayed by the formation of the RNase T1-resistant oligonucleotide VPg-pUUAAAACAGp or by fingerprint analysis of the in vitro-synthesized RNA. Incubation of preformed VPg-pUpU with unlabeled nucleoside triphosphates resulted in the formation of VPg-pUUAAAACAGp. This reaction, which appeared to be an elongation of VPg-pUpU, was stimulated by the addition of a soluble fraction (S-10) obtained from uninfected HeLa cells. Preformed VPg-pU could be chased into VPg-pUpU in the presence of UTP. Our data are consistent with a model that VPg-pU can function as a primer for poliovirus plus-strand RNA synthesis in the membranous replication complex and that the elongation reaction may be stimulated by a host cellular factor.

MeSH Terms
Cell Membrane/physiology Cell-Free System HeLa Cells Humans Oligoribonucleotides/analysis Poliovirus/physiology RNA, Viral/biosynthesis Viral Core Proteins Viral Proteins/analysis Virus Replication
Chemicals
Oligoribonucleotides RNA, Viral VPg-pUpU Viral Core Proteins Viral Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Takeda N
Kuhn R J
Yang C F
Takegami T
Wimmer E
References (51)
51 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. Interconversion of linear and circular forms of double-stranded RNA of encephalomyocarditis virus.
    Virology. 1979 Mar;93(2):574-7 PMID: 222058
  3. Characterization of poliovirus-specific structures associated with cytoplasmic membranes.
    Virology. 1970 Sep;42(1):112-22 PMID: 4318975
  4. Kinetics of appearance of the products of poliovirus-induced RNA polymerase.
    Virology. 1970 Dec;42(4):999-1006 PMID: 4321316
  5. Chemical and enzymatic characteristics of cytoplasmic membranes of poliovirus-infected HeLa cells.
    Virology. 1972 Jan;47(1):30-8 PMID: 4333575
  6. In vivo and in vitro synthesis of human rhinovirus type 2 ribonucleic acid.
    J Virol. 1972 Jul;10(1):93-8 PMID: 4339200
  7. Polyadenylic acid at the 3'-terminus of poliovirus RNA.
    Proc Natl Acad Sci U S A. 1972 Jul;69(7):1877-82 PMID: 4340162
  8. The formation of poliovirus particles in association with the RNA replication complexes.
    J Gen Virol. 1973 Oct;21:99-108 PMID: 4357379
  9. Poly(U) in poliovirus minus RNA is 5'-terminal.
    Biochem Biophys Res Commun. 1974 Dec 23;61(4):1101-9 PMID: 4376003
  10. Replication of picornaviruses. I. Evidence from in vitro RNA synthesis that poly(A) of the poliovirus genome is genetically coded.
    J Virol. 1975 Dec;16(6):1512-7 PMID: 172656
  11. "Fingerprinting" high molecular weight RNA by two-dimensional gel electrophoresis: application to poliovirus RNA.
    Nucleic Acids Res. 1976 Jul;3(7):1647-58 PMID: 184435
  12. A protein covalently linked to poliovirus genome RNA.
    Proc Natl Acad Sci U S A. 1977 Jan;74(1):59-63 PMID: 189316
  13. A new method for sequencing DNA.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
  14. Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus.
    Proc Natl Acad Sci U S A. 1977 Mar;74(3):961-5 PMID: 191841
  15. The location of the polio genome protein in viral RNAs and its implication for RNA synthesis.
    Nature. 1977 Jul 21;268(5617):208-13 PMID: 196204
  16. Mapping adenines, guanines, and pyrimidines in RNA.
    Nucleic Acids Res. 1977 Aug;4(8):2527-38 PMID: 409999
  17. Poliovirus-specific primer-dependent RNA polymerase able to copy poly(A).
    Proc Natl Acad Sci U S A. 1977 Sep;74(9):3677-80 PMID: 198796
  18. The 5'-terminal structures of poliovirion RNA and poliovirus mRNA differ only in the genome-linked protein VPg.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5345-9 PMID: 202952
  19. Protein is linked to the 5' end of poliovirus RNA by a phosphodiester linkage to tyrosine.
    J Biol Chem. 1978 Aug 10;253(15):5263-6 PMID: 209034
  20. Identification of a protein linked to nascent poliovirus RNA and to the polyuridylic acid of negative-strand RNA.
    J Virol. 1978 Aug;27(2):357-65 PMID: 211265
  21. O4-(5'-uridylyl)tyrosine is the bond between the genome-linked protein and the RNA of poliovirus.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4868-72 PMID: 217003
  22. Nucleotide sequences adjacent to the proteins covalently linked to the cowpea mosaic virus genome. Sequence determination after labelling in vitro using RNA ligase.
    Eur J Biochem. 1979 Nov 1;101(1):45-9 PMID: 116852
  23. Sequence studies of poliovirus RNA. IV. Nucleotide sequence complexities of poliovirus type 1, type 2 and two type 1 defective interfering particles RNAs, and fingerprint of the poliovirus type 3 genome.
    J Gen Virol. 1979 Aug;44(2):311-22 PMID: 230285
  24. Kinetics and location of poliovirus macromolecular synthesis in correlation to virus-induced cytopathology.
    Virology. 1980 Jan 30;100(2):390-9 PMID: 6243433
  25. Dependence of the activity of the poliovirus replicase on the host cell protein.
    Cell. 1980 Feb;19(2):423-9 PMID: 6244109
  26. Isolation of a soluble and template-dependent poliovirus RNA polymerase that copies virion RNA in vitro.
    J Virol. 1979 Oct;32(1):155-61 PMID: 232168
  27. Palindrome-like dimers of double-stranded RNA of encephalomyocarditis virus.
    Virology. 1980 Apr 30;102(2):339-48 PMID: 6245513
  28. A micromethod for detailed characterization of high molecular weight RNA.
    Methods Enzymol. 1980;65(1):680-7 PMID: 6154875
  29. Heterogeneity of the 3' end of minus-strand RNA in the poliovirus replicative form.
    J Virol. 1980 Nov;36(2):387-94 PMID: 6253664
  30. The structure of poliovirus replicative form.
    Nucleic Acids Res. 1980 Mar 25;8(6):1217-29 PMID: 6253931
  31. Phy M: an RNase activity specific for U and A residues useful in RNA sequence analysis.
    Nucleic Acids Res. 1980 Jul 25;8(14):3133-42 PMID: 6160466
  32. Comparison of replication complexes synthesizing poliovirus RNA.
    Virology. 1981 May;111(1):33-46 PMID: 6263009
  33. Primary structure, gene organization and polypeptide expression of poliovirus RNA.
    Nature. 1981 Jun 18;291(5816):547-53 PMID: 6264310
  34. Poliovirus replication proteins: RNA sequence encoding P3-1b and the sites of proteolytic processing.
    Proc Natl Acad Sci U S A. 1981 Jun;78(6):3464-8 PMID: 6267593
  35. Genome-linked proteins of viruses.
    Cell. 1982 Feb;28(2):199-201 PMID: 7060125
  36. A membrane-associated precursor to poliovirus VPg identified by immunoprecipitation with antibodies directed against a synthetic heptapeptide.
    Cell. 1982 Feb;28(2):405-12 PMID: 6277514
  37. Genome-length copies of poliovirion RNA are synthesized in vitro by the poliovirus RNA-dependent RNA polymerase.
    J Biol Chem. 1982 Apr 25;257(8):4610-7 PMID: 6279640
  38. 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
  39. In vitro copying of viral positive strand RNA by poliovirus replicase. Characterization of the reaction and its products.
    J Biol Chem. 1982 Oct 25;257(20):12359-66 PMID: 6288719
  40. Membrane fractions active in poliovirus RNA replication contain VPg precursor polypeptides.
    Virology. 1983 Jul 15;128(1):33-47 PMID: 6308897
  41. Intracellular distribution of poliovirus proteins and the induction of virus-specific cytoplasmic structures.
    Virology. 1983 Nov;131(1):39-48 PMID: 6316654
  42. Membrane-dependent uridylylation of the genome-linked protein VPg of poliovirus.
    Proc Natl Acad Sci U S A. 1983 Dec;80(24):7447-51 PMID: 6324172
  43. Systematic nomenclature of picornavirus proteins.
    J Virol. 1984 Jun;50(3):957-9 PMID: 6726891
  44. Association of poliovirus proteins with the endoplasmic reticulum.
    J Virol. 1984 Dec;52(3):777-83 PMID: 6092710
  45. Antibody to poliovirus genome-linked protein (VPg) precipitates in vitro synthesized RNA attached to VPg-precursor polypeptide(s).
    Virus Res. 1984;1(2):89-100 PMID: 6532002
  46. The host protein required for in vitro replication of poliovirus is a protein kinase that phosphorylates eukaryotic initiation factor-2.
    Cell. 1985 Apr;40(4):913-21 PMID: 2985268
  47. Poliovirus RNA-dependent RNA polymerase and host cell protein synthesize product RNA twice the size of poliovirion RNA in vitro.
    J Virol. 1985 May;54(2):256-64 PMID: 2985794
  48. Poliovirus replicase stimulation by terminal uridylyl transferase.
    J Biol Chem. 1985 Jun 25;260(12):7628-35 PMID: 2987262
  49. Purification of a terminal uridylyltransferase that acts as host factor in the in vitro poliovirus replicase reaction.
    Proc Natl Acad Sci U S A. 1986 Jan;83(2):221-5 PMID: 2417240
  50. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  51. The role of cytoplasmic membranes in poliovirus biosynthesis.
    Virology. 1970 Sep;42(1):100-11 PMID: 4318974
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.60.1.43-53.1986
Published
1986-10-00
Pages
43-53
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC253900
Subset
IM
Grants
NIAID NIH HHS · AI 15122 · United States
NCI NIH HHS · CA 28146 · United States
External Links
PubMed source DOI: 10.1128/JVI.60.1.43-53.1986 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2