Home LiteratureArticle Details
PMID: 11044081 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Genetic and biochemical studies of poliovirus cis-acting replication element cre in relation to VPg uridylylation.

Journal of virology ·Vol. 74 ·No. 22 ·2000-11-00 ·Pages 10371-80

Rieder E, Paul AV, Kim DW, van Boom JH, Wimmer E

Abstract

In addition to highly conserved stem-loop structures located in the 5'- and 3'-nontranslated regions, genome replication of picornaviruses requires cis-acting RNA elements located in the coding region (termed cre) (K. L. McKnight and S. M. Lemon, J. Virol. 70:1941-1952, 1996; P. E. Lobert, N. Escriou, J. Ruelle, and T. Michiels, Proc. Natl. Acad. Sci. USA 96:11560-11565, 1999; I. Goodfellow, Y. Chaudhry, A. Richardson, J. Meredith, J. W. Almond, W. Barclay, and D. J. Evans, J. Virol. 74:4590-4600, 2000). cre elements appear to be essential for minus-strand RNA synthesis by an as-yet-unknown mechanism. We have discovered that the cre element of poliovirus (mapping to the 2C coding region of poliovirus type 1; nucleotides 4444 to 4505 in 2C), which is homologous to the cre element of poliovirus type 3, is preferentially used as a template for the in vitro uridylylation of VPg catalyzed by 3D(pol) in a reaction that is greatly stimulated by 3CD(pro) (A. V. Paul, E. Rieder, D. W. Kim, J. H. van Boom, and E. Wimmer, J. Virol. 74:10359-10370, 2000). Here we report a direct correlation between mutations that eliminate, or severely reduce, the in vitro VPg-uridylylation reaction and produce replication phenotypes in vivo. None of the genetic changes significantly influenced translation or polyprotein processing. A substitution mapping to the first A (A4472C) of a conserved AAACA sequence in the loop of PV-cre(2C) eliminated the ability of the cre RNA to serve as template for VPg uridylylation and abolished RNA infectivity. Mutagenesis of the second A (A4473C; AAACA) severely reduced the yield of VPgpUpU and RNA infectivity was restored only after reversion to the wild-type sequence. The effect of substitution of the third A (A4474G; AAACA) was less severe but reduced both VPg uridylylation and virus yield. Disruption of base pairing within the upper stem region of PV-cre(2C) also affected uridylylation of VPg. Virus derived from transcripts containing mutations in the stem was either viable or quasi-infectious.

MeSH Terms
Base Sequence HeLa Cells Humans Molecular Sequence Data Mutation Nucleic Acid Conformation Phenotype Plasmids/genetics Poliovirus/chemistry,genetics,physiology RNA, Viral/chemistry,genetics,metabolism Transfection Uridine Monophosphate/metabolism Viral Core Proteins/metabolism Viral Proteins/genetics,metabolism Virus Replication/genetics
Chemicals
RNA, Viral Viral Core Proteins Viral Proteins Uridine Monophosphate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Rieder E
Department of Molecular Genetics and Microbiology, State University of New York at Stony Brook, Stony Brook, New York 11794, USA.
Paul A V
Kim D W
van Boom J H
Wimmer E
References (43)
43 references, click to expand
  1. Identification of a cis-acting replication element within the poliovirus coding region.
    J Virol. 2000 May;74(10):4590-600 PMID: 10775595
  2. 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
  3. Identification of an RNA hairpin in poliovirus RNA that serves as the primary template in the in vitro uridylylation of VPg.
    J Virol. 2000 Nov;74(22):10359-70 PMID: 11044080
  4. Polyadenylic acid at the 3'-terminus of poliovirus RNA.
    Proc Natl Acad Sci U S A. 1972 Jul;69(7):1877-82 PMID: 4340162
  5. Poly(U) in poliovirus minus RNA is 5'-terminal.
    Biochem Biophys Res Commun. 1974 Dec 23;61(4):1101-9 PMID: 4376003
  6. 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
  7. A protein covalently linked to poliovirus genome RNA.
    Proc Natl Acad Sci U S A. 1977 Jan;74(1):59-63 PMID: 189316
  8. Polioviruses containing picornavirus type 1 and/or type 2 internal ribosomal entry site elements: genetic hybrids and the expression of a foreign gene.
    Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1406-10 PMID: 7509072
  9. Genetics of poliovirus.
    Annu Rev Genet. 1993;27:353-436 PMID: 8122908
  10. The 5'-untranslated regions of picornavirus RNAs contain independent functional domains essential for RNA replication and translation.
    J Virol. 1994 Jul;68(7):4384-91 PMID: 8207812
  11. 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
  12. Mouse neurovirulence determinants of poliovirus type 1 strain LS-a map to the coding regions of capsid protein VP1 and proteinase 2Apro.
    J Virol. 1994 Nov;68(11):7507-15 PMID: 7933134
  13. Interaction between the 5'-terminal cloverleaf and 3AB/3CDpro of poliovirus is essential for RNA replication.
    J Virol. 1995 Jun;69(6):3658-67 PMID: 7745714
  14. Host cell proteins binding to domain IV of the 5' noncoding region of poliovirus RNA.
    J Virol. 1995 Jul;69(7):4381-9 PMID: 7769700
  15. The 3' untranslated region of picornavirus RNA: features required for efficient genome replication.
    J Virol. 1995 Dec;69(12):7835-44 PMID: 7494295
  16. Molecular dissection of the multifunctional poliovirus RNA-binding protein 3AB.
    RNA. 1995 Nov;1(9):892-904 PMID: 8548654
  17. Capsid coding sequence is required for efficient replication of human rhinovirus 14 RNA.
    J Virol. 1996 Mar;70(3):1941-52 PMID: 8627720
  18. Complete nucleotide sequence of enterovirus 71 is distinct from poliovirus.
    Virus Res. 1995 Dec;39(2-3):195-205 PMID: 8837884
  19. Cis-element, oriR, involved in the initiation of (-) strand poliovirus RNA: a quasi-globular multi-domain RNA structure maintained by tertiary ('kissing') interactions.
    EMBO J. 1996 Oct 1;15(19):5428-36 PMID: 8895586
  20. Kissing of the two predominant hairpin loops in the coxsackie B virus 3' untranslated region is the essential structural feature of the origin of replication required for negative-strand RNA synthesis.
    J Virol. 1997 Jan;71(1):686-96 PMID: 8985400
  21. Two functional complexes formed by KH domain containing proteins with the 5' noncoding region of poliovirus RNA.
    RNA. 1997 Aug;3(8):882-92 PMID: 9257647
  22. Poly (rC) binding protein 2 forms a ternary complex with the 5'-terminal sequences of poliovirus RNA and the viral 3CD proteinase.
    RNA. 1997 Oct;3(10):1124-34 PMID: 9326487
  23. Replication-competent picornaviruses with complete genomic RNA 3' noncoding region deletions.
    J Virol. 1997 Nov;71(11):8868-74 PMID: 9343250
  24. Protein-primed RNA synthesis by purified poliovirus RNA polymerase.
    Nature. 1998 May 21;393(6682):280-4 PMID: 9607767
  25. Switch from translation to RNA replication in a positive-stranded RNA virus.
    Genes Dev. 1998 Aug 1;12(15):2293-304 PMID: 9694795
  26. The rhinovirus type 14 genome contains an internally located RNA structure that is required for viral replication.
    RNA. 1998 Dec;4(12):1569-84 PMID: 9848654
  27. Characterization of the nucleoside triphosphatase activity of poliovirus protein 2C reveals a mechanism by which guanidine inhibits poliovirus replication.
    J Biol Chem. 1999 Mar 12;274(11):6992-7001 PMID: 10066753
  28. A coding RNA sequence acts as a replication signal in cardioviruses.
    Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11560-5 PMID: 10500216
  29. Paradoxes of the replication of picornaviral genomes.
    Virus Res. 1999 Aug;62(2):129-47 PMID: 10507323
  30. A cysteine-rich motif in poliovirus protein 2C(ATPase) is involved in RNA replication and binds zinc in vitro.
    J Virol. 2000 Jan;74(1):334-43 PMID: 10590122
  31. 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
  32. 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
  33. 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
  34. Poliovirus single-stranded RNA and double-stranded RNA: differential infectivity in enucleate cells.
    J Virol. 1978 Sep;27(3):582-6 PMID: 212582
  35. 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
  36. 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
  37. Production of guanidine-resistant and -dependent poliovirus mutants from cloned cDNA: mutations in polypeptide 2C are directly responsible for altered guanidine sensitivity.
    J Virol. 1986 Nov;60(2):793-6 PMID: 3022012
  38. Protein 3CD is the major poliovirus proteinase responsible for cleavage of the P1 capsid precursor.
    Virology. 1988 Sep;166(1):265-70 PMID: 2842953
  39. A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA.
    Cell. 1990 Oct 19;63(2):369-80 PMID: 2170027
  40. Protein-priming of DNA replication.
    Annu Rev Biochem. 1991;60:39-71 PMID: 1883199
  41. Cell-free, de novo synthesis of poliovirus.
    Science. 1991 Dec 13;254(5038):1647-51 PMID: 1661029
  42. Functional and genetic plasticities of the poliovirus genome: quasi-infectious RNAs modified in the 5'-untranslated region yield a variety of pseudorevertants.
    J Virol. 1993 Oct;67(10):6309-16 PMID: 8396686
  43. Studies on the attenuation phenotype of polio vaccines: poliovirus RNA polymerase derived from Sabin type 1 sequence is temperature sensitive in the uridylylation of VPg.
    Virology. 2000 Jun 20;272(1):72-84 PMID: 10873750
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.22.10371-10380.2000
Published
2000-11-00
Pages
10371-80
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC110911
Subset
IM
Grants
NIAID NIH HHS · R37 AI015122 · United States
NIAID NIH HHS · 5R37AI15122 · United States
External Links
PubMed source DOI: 10.1128/jvi.74.22.10371-10380.2000 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