Poliovirus RNA-Dependent RNA Polymerase Synthesizes Full-Length Copies of Poliovirion RNA, Cellular mRNA, and Several Plant Virus RNAs In Vitro

  title={Poliovirus RNA-Dependent RNA Polymerase Synthesizes Full-Length Copies of Poliovirion RNA, Cellular mRNA, and Several Plant Virus RNAs In Vitro},
  author={D M Tuschall and Ernest Hiebert and James B. Flanegan},
  journal={Journal of Virology},
  pages={209 - 216}
The poliovirus RNA-dependent RNA polymerase was active on synthetic homopolymeric RNA templates as well as on every natural RNA tested. The polymerase copied polyadenylate· oligouridylate [oligo(U)], polycytidylate · oligoinosinate, and polyinosinate· oligocytidylate templates to about the same extent. The observed activity on polyuridylate· oligoadenylate was about fourfold less. Full-length copies of both poliovirion RNA and a wide variety of other polyadenylated RNAs were synthesized by the… 

Poliovirus RNA-dependent RNA polymerase and host cell protein synthesize product RNA twice the size of poliovirion RNA in vitro

The results from this study indicate that the labeled negative-strand product RNA synthesized in vitro was covalently linked to the positive-Strand template RNA.

Synthesis of plus- and minus-strand RNA from poliovirion RNA template in vitro

The poliovirus RNA polymerase, 3Dpol, was used to synthesize RNA in vitro in the presence of a host factor preparation from uninfected HeLa cells and poliovirion RNA as the template, demonstrating that host factor-mediated polymerase products contain newly synthesized plus-strand sequences as well as the expected minus-Strand sequences.

Direct measurement of the poliovirus RNA polymerase error frequency in vitro

The fidelity of RNA replication by the poliovirus-RNA-dependent RNA polymerase was examined by copying homopolymeric RNA templates in vitro and there were no significant differences among the error frequencies obtained with different noncomplementary nucleotide substrates on a given template.

Poliovirus RNA replication.

Historically, the experimental approach initially utilized to analyze the poliovirus RNA replication reaction was enzymological; efforts were made to isolate and purify an RNA-dependent RNA polymerase activity from virus-infected cells, but this approach yielded little information about the mechanism of RNA replication.

Anti-VPg antibody precipitation of product RNA synthesized in vitro by the poliovirus polymerase and host factor is mediated by VPg on the poliovirion RNA template

The results support the previous evidence that a covalent linkage exists between the labeled negative-strand product RNA and the VPg-linked template RNA and suggest that the purified polymerase and host factor initiated RNA synthesis in vitro in the absence of VPg or aVPg-precursor protein.

Purification and properties of poliovirus RNA polymerase expressed in Escherichia coli

A cDNA clone encoding the RNA polymerase of poliovirus has been expressed in Escherichia coli under the transcriptional control of a T7 bacteriophage promoter, and the enzyme exhibits poly(A)-dependent oligo(U)-primed poly(U) polymerase activity as well as RNA polymer enzyme activity.

Poliovirus protein 3AB forms a complex with and stimulates the activity of the viral RNA polymerase, 3Dpol

A mechanism for bringing together 3AB, 3Dpol (or its precursor 3CD), and viral RNA in host cell membranous vesicles in which all viral RNA synthesis occurs is suggested.

Synchronous replication of poliovirus RNA: initiation of negative-strand RNA synthesis requires the guanidine-inhibited activity of protein 2C

It is suggested that guanidine may block an irreversible structural maturation of protein 2C and/or RNA replication complexes necessary for the initiation of RNA replication.

ATP is required for initiation of poliovirus RNA synthesis in vitro: demonstration of tyrosine-phosphate linkage between in vitro-synthesized RNA and genome-linked protein

The poly(U) synthesized in a host factor-stimulated reaction was shown to be attached to VPg precursor polypeptide(s) via a tyrosine-phosphate bond as found in poliovirion VPg-RNA.

Partial purification and characterization of Cucumber necrosis virus and Tomato bushy stunt virus RNA-dependent RNA polymerases: similarities and differences in template usage between tombusvirus and carmovirus RNA-dependent RNA polymerases.

Generation of full-length, complementary RNA by the tombusvirus RdRp suggests that it can correctly and efficiently recognize the heterologous TCV-specific promoters.



Genome-length copies of poliovirion RNA are synthesized in vitro by the poliovirus RNA-dependent RNA polymerase.

Polyadenylic acid on poliovirus RNA. II. poly(A) on intracellular RNAs

Investigation of the kinetics of poly(A) appearance in the replicative intermediate and in finished 35S molecules indicated that poly (A) is the last portion of the 35S RNA to be synthesized, which indicates that poliovirus RNA is synthesized 5' leads to 3' like other RNA's.

Isolation of a soluble and template-dependent poliovirus RNA polymerase that copies virion RNA in vitro

A soluble RNA-dependent RNA polymerase was isolated from poliovirus-infected HeLa cells and was shown to copy poliov virus RNA in vitro, suggesting that the product RNA was complementary to the virion RNA template.

Poliovirus-specific primer-dependent RNA polymerase able to copy poly(A).

  • J. FlaneganD. Baltimore
  • Biology
    Proceedings of the National Academy of Sciences of the United States of America
  • 1977
Analysis of the polymerase by glycerol gradient centrifugation showed that the majority of the activity sedimented at about 4 S, indicating that it was no longer complexed with high-molecular-weight RNA or cellular membranes.

Identification of poliovirus polypeptide P63 as a soluble RNA-dependent RNA polymerase

The viral polypeptide, p63, is an RNA-dependent RNA polymerase that can copy poliovirion RNA when oligouridylic acid is used as a primer.

Poliovirus Polyuridylic Acid Polymerase and RNA Replicase Have the Same Viral Polypeptide

A poliovirus-specific polyuridylic acid [poly(U)] polymerase that copies a polyadenylic acid template complexed to an oligouridylic acid primer was isolated from the membrane fraction of infected

Poliovirus replicase: a soluble enzyme able to initiate copying of poliovirus RNA.

Although less pure replicase fractions copy a variety of RNAs, purer fractions respond better to poliovirus RNA than to other viral RNAs.

Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus.

Digests of replicative intermediate RNA contained sufficient protein-pUp to suggest that this structure is at the 5'-end of most nascent poliovirion RNA molecules, suggesting that a protein-nucleotide structure acts as a primer for initiating synthesis of poliovirus RNA.