• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. Dec 1975; 16(6): 1512–1517.
PMCID: PMC355760

Replication of picornaviruses. I. Evidence from in vitro RNA synthesis that poly(A) of the poliovirus genome is genetically coded.


A crude replication complex has been isolated from poliovirus-infected HeLa cells and used for synthesis of poliovirus replicative intermediate (RI) RNA, replicative form (RF) RNA, and single-stranded (SS) RNA in vitro. All three classes of virus-specific RNA synthesized in vitro are shown to contain poly(A). Poly(A) of RF and of SS RNA [RF-poly(A) and SS-poly(A)] has a chain length (50 to 70 nucleotides) that is shorter than that of poly(A) of in vivo-synthesized RNAs. Poly(A) of RI [RI-poly(A),] however, is at least 200 nucleotides long and, therefore, larger than poly(A) of RI isolated from HeLa cells 4 h after infection. The crude membrane-bound replication complex contains a terminal adenylate transferase activity that is stimulated by Mn2+ and the addition of an (Ap)2AOH primer. This transferase activity is found also in extracts of mock-infected cells. Partial purificaiton of the replication complex in a stepwise sucrose gradient, in which the viral replicase is associated with the smooth cytoplasmic membrane fraction, does not remove the terminal transferase. However, when the partially purified replication complex is treated with deoxycholate and sedimented through a sucrose gradient, a soluble replication complex can be isolated that is free from terminal adenylate transferase. This soluble replication complex was found to synthesize viral RNA-linked poly(A) longer in chain length than that synthesized by the crude replication complex. Taking into account the 5'-terminal poly(U) in poliovirus minus strands, our data suggest that polyadenylation of poliovirus RNA occurs by transcription and not by end addition. When compared to other viral systems, poliovirus and, probably, all picornaviruses appear to be unique in that the poly(A) of their genome is genetically coded.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.3M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adesnik M, Darnell JE. Biogenesis and characterization of histone messenger RNA in HeLa cells. J Mol Biol. 1972 Jun 28;67(3):397–406. [PubMed]
  • Armstrong JA, Edmonds M, Nakazato H, Phillips BA, Vaughn MH. Polyadenylic acid sequences in the virion RNA of poliovirus and Eastern Equine Encephalitis virus. Science. 1972 May 5;176(4034):526–528. [PubMed]
  • BALTIMORE D. IN VITRO SYNTHESIS OF VIRAL RNA BY THE POLIOVIRUS RNA POLYMERASE. Proc Natl Acad Sci U S A. 1964 Mar;51:450–456. [PMC free article] [PubMed]
  • Baltimore D. Structure of the poliovirus replicative intermediate RNA. J Mol Biol. 1968 Mar 14;32(2):359–368. [PubMed]
  • Baltimore D. Expression of animal virus genomes. Bacteriol Rev. 1971 Sep;35(3):235–241. [PMC free article] [PubMed]
  • Baltimore D, Girard M. An intermediate in the synthesis of poliovirus RNA. Proc Natl Acad Sci U S A. 1966 Aug;56(2):741–748. [PMC free article] [PubMed]
  • Baltimore D, Huang AS, Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus, II. An RNA polymerase in the virion. Proc Natl Acad Sci U S A. 1970 Jun;66(2):572–576. [PMC free article] [PubMed]
  • Banerjee AK, Moyer SA, Rhodes DP. Studies on the in vitro adenylation of RNA by vesicular stomatitis virus. Virology. 1974 Oct;61(2):547–558. [PubMed]
  • BEERS RF., Jr Hydrolysis of polyadenylic acid by pancreatic ribonuclease. J Biol Chem. 1960 Aug;235:2393–2398. [PubMed]
  • Bishop JM, Koch G. Infectious replicative intermediate of poliovirus: purification and characterization. Virology. 1969 Apr;37(4):521–534. [PubMed]
  • Brawerman G. Eukaryotic messenger RNA. Annu Rev Biochem. 1974;43(0):621–642. [PubMed]
  • Brown M, Dorson JW, Bollum FJ. Terminal riboadenylate transferase: a poly A polymerase in purified vaccinia virus. J Virol. 1973 Aug;12(2):203–208. [PMC free article] [PubMed]
  • Caliguiri LA. Analysis of RNA associated with the poliovirus RNA replication complexes. Virology. 1974 Apr;58(2):526–535. [PubMed]
  • Caliguiri LA, Compans RW. The formation of poliovirus particles in association with the RNA replication complexes. J Gen Virol. 1973 Oct;21:99–108. [PubMed]
  • Caliguiri LA, Mosser AG. Proteins associated with the poliovirus RNA replication complex. Virology. 1971 Nov;46(2):375–386. [PubMed]
  • Caliguiri LA, Tamm I. The role of cytoplasmic membranes in poliovirus biosynthesis. Virology. 1970 Sep;42(1):100–111. [PubMed]
  • Ehrenfeld E. Polyadenylation of vesicular stomatitis virus mRNA. J Virol. 1974 May;13(5):1055–1060. [PMC free article] [PubMed]
  • Ehrenfeld E, Maizel JV, Summers DF. Soluble RNA polymerase complex from poliovirus-infected HeLa cells. Virology. 1970 Apr;40(4):840–846. [PubMed]
  • Girard M. In vitro synthesis of poliovirus ribonucleic acid: role of the replicative intermediate. J Virol. 1969 Apr;3(4):376–384. [PMC free article] [PubMed]
  • Katz L, Penman S. The solvent denaturation of double-stranded RNA from poliovirus infected HeLa cells. Biochem Biophys Res Commun. 1966 May 25;23(4):557–560. [PubMed]
  • Lewin B. Units of transcription and translation: the relationship between heterogeneous nuclear RNA and messenger RNA. Cell. 1975 Jan;4(1):11–20. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • McDonnell JP, Levintow L. Kinetics of appearance of the products of poliovirus-induced RNA polymerase. Virology. 1970 Dec;42(4):999–1006. [PubMed]
  • Marshall S, Gillespie D. Poly U tracts absent from viral RNA. Nat New Biol. 1972 Nov 8;240(97):43–45. [PubMed]
  • Milcarek C, Price R, Penman S. The metabolism of a poly(A) minus mRNA fraction in HeLa cells. Cell. 1974 Sep;3(1):1–10. [PubMed]
  • Moss B, Rosenblum EN, Paoletti E. Polyadenylate polymerase from vaccinia virions. Nat New Biol. 1973 Sep 12;245(141):59–63. [PubMed]
  • Nemer M, Graham M, Dubroff LM. Co-existence of non-histone messenger RNA species lacking and containing polyadenylic acid in sea urchin embryos. J Mol Biol. 1974 Nov 5;89(3):435–454. [PubMed]
  • Nevins JR, Joklik WK. Poly (A) sequences of vaccinia virus messenger RNA: nature, mode of addition and function during translation in vitra and in vivo. Virology. 1975 Jan;63(1):1–14. [PubMed]
  • Oberg B, Philipson L. Replicative structures of poliovirus RNA in vivo. J Mol Biol. 1971 Jun 28;58(3):725–737. [PubMed]
  • Olivera BM, Lehman IR. Enzymic joining of polynucleotides. 3. The polydeoxyadenylate-polydeoxythymidylate homopolymer pair. J Mol Biol. 1968 Sep 14;36(2):261–274. [PubMed]
  • Penman S, Rosbash M, Penman M. Messenger and heterogeneous nuclear RNA in HeLa cells: differential inhibition by cordycepin. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1878–1885. [PMC free article] [PubMed]
  • Roeder RG. Multiple forms of deoxyribonucleic acid-dependent ribonucleic acid polymerase in Xenopus laevis. Isolation and partial characterization. J Biol Chem. 1974 Jan 10;249(1):241–248. [PubMed]
  • Roeder RG, Rutter WJ. Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969 Oct 18;224(5216):234–237. [PubMed]
  • Roumiantzeff M, Maizel JV, Jr, Summers DF. Comparison of polysomal structures of uninfected and poliovirus infected HeLa cells. Virology. 1971 May;44(2):239–248. [PubMed]
  • Shatkin AJ. Animal RNA viruses: genome structure and function. Annu Rev Biochem. 1974;43(0):643–665. [PubMed]
  • Sheldon R, Jurale C, Kates J. Detection of polyadenylic acid sequences in viral and eukaryotic RNA(polu(U)-cellulose columns-poly(U) filters-fiberglass-HeLa cells-bacteriophage T4). Proc Natl Acad Sci U S A. 1972 Feb;69(2):417–421. [PMC free article] [PubMed]
  • Sheldon R, Kates J. Mechanism of poly(A) synthesis by vaccinia virus. J Virol. 1974 Aug;14(2):214–224. [PMC free article] [PubMed]
  • Soria M, Little SP, Huang AS. Characterization of vesicular stomatitis virus nucleocapsids. I. Complementary 40 S RNA molecules in nucleocapsids. Virology. 1974 Sep;61(1):270–280. [PubMed]
  • Spector DH, Baltimore D. Requirement of 3'-terminal poly(adenylic acid) for the infectivity of poliovirus RNA. Proc Natl Acad Sci U S A. 1974 Aug;71(8):2983–2987. [PMC free article] [PubMed]
  • Spector DH, Baltimore D. Polyadenylic acid on poliovirus RNA. III. In vitro addition of polyadenylic acid to poliovirus RNAs. J Virol. 1975 Jun;15(6):1432–1439. [PMC free article] [PubMed]
  • Stoltzfus CM, Shatkin AJ, Banerjee AK. Absence of polyadenylic acid from reovirus messenger ribonucleic acid. J Biol Chem. 1973 Dec 10;248(23):7993–7998. [PubMed]
  • Tsiapalis CM, Dorson JW, De Sante DM, Bollum FJ. Terminal riboadenylate transferase: a polyadenylate polymerase from calf thymus gland. Biochem Biophys Res Commun. 1973 Feb 5;50(3):737–743. [PubMed]
  • Villarreal LP, Holland JJ. Synthesis of poly(A) in vitro by purified virions of vesicular stomatitis virus. Nat New Biol. 1973 Nov 7;246(149):17–19. [PubMed]
  • Yogo Y, Teng MH, Wimmer E. Poly(U) in poliovirus minus RNA is 5'-terminal. Biochem Biophys Res Commun. 1974 Dec 23;61(4):1101–1109. [PubMed]
  • Yogo Y, Wimmer E. Polyadenylic acid at the 3'-terminus of poliovirus RNA. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1877–1882. [PMC free article] [PubMed]
  • Yogo Y, Wimmer E. Poly (A) and poly (U) in poliovirus double stranded RNA. Nat New Biol. 1973 Apr 11;242(119):171–174. [PubMed]
  • Yogo Y, Wimmer E. Sequence studies of poliovirus RNA. III. Polyuridylic acid and polyadenylic acid as components of the purified poliovirus replicative intermediate. J Mol Biol. 1975 Mar 5;92(3):467–477. [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem Compound links
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...