Construction and properties of intertypic poliovirus recombinants: First approximation mapping of the major determinants of neurovirulence

Agol, Vadim I.; Grachev, V. T.; Drozdov, S. G.; Kolesnikova, Marina S.; Kozlov, V.G.; Ralph, N. M.; Romanova, Lyudmila I.; Tolskaya, Elena A.; Tyufanov, A. V.; Viktorova, Ekaterina G.

doi:10.1016/0042-6822(84)90246-0

Cited by 50 publications

(22 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Alternatively, RNA pulse labelling of 30 rain from 5 to 5-5 h post-infection was done. In this case, dactinomycin (2.5 Ixg/ml) was added only 15 rain prior to the addition of [3H]aridine (Agol et aL, 1984). …”

Section: Methodsmentioning

confidence: 99%

Analysis of the functional significance of amino acid residues in the putative NTP-binding pattern of the poliovirus 2C protein

Teterina

Kean

Gorbalenya

et al. 1992

Journal of General Virology

View full text Add to dashboard Cite

The amino acid sequence of the poliovirus 2C protein contains two highly conserved stretches, GSPGTGKS t36 and MDD 177, which correspond to the consensus 'A' and 'B' motifs (GXXXXGKS/T and DD/E, respectively) found in nucleoside triphosphatebinding proteins. To assess the functional importance of these amino acid sequences, we changed conserved and non-conserved amino acids. The replacement of the non-conserved Thr 133 residue with Ser or Ala did not markedly change the virus phenotype. Similarly, replacement of the non-conserved Pro TM residue by Ala did not abolish virus viability, but changes of this residue to Thr or Asn were not tolerated. No viable mutant could be isolated after transfection of cultured cells with transcripts mutated at the conserved Lys 135, Ser 136 or Asp ~77 residues. However, true revertants were selected from Arg 135 and Ser ~35 mutants, from Glu 177 and Gly 177 mutants, and from Ala 136 mutants. Thr ~ 36 mutants not only gave rise to true revertants, but also to two independent isolates of a suppressor mutant, Asn14°-~Tyr. All the lethal mutations resulted in severe inhibition of viral RNA synthesis in vivo, although no translational deficiency was detected in a cell-free system. This is the first direct evidence for the functional significance of the nucleoside triphosphatebinding pattern in the poliovirus 2C protein.

show abstract

Section: Methodsmentioning

confidence: 99%

Analysis of the functional significance of amino acid residues in the putative NTP-binding pattern of the poliovirus 2C protein

Teterina

Kean

Gorbalenya

et al. 1992

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…The inability of virus to spread to tissues such as kidney, heart, and lung may be a consequence of host range restrictions that are independent of receptor binding. Determinants of the attenuation phenotypes for the Sabin strains of poliovirus have been mapped to the 5Ј-proximal portion of genomic RNAs of all three strains (4,8,14,18,23,24,33,39,54). Shiroki et al induced a host-range phenotype of poliovirus by introducing mutations in the internal ribosomal entry site of the poliovirus genome (47).…”

mentioning

confidence: 99%

Cell-Dependent Role for the Poliovirus 3′ Noncoding Region in Positive-Strand RNA Synthesis

Brown

Kauder

Cornell

et al. 2004

J Virol

View full text Add to dashboard Cite

We previously reported the isolation of a mutant poliovirus lacking the entire genomic RNA 3 noncoding region. Infection of HeLa cell monolayers with this deletion mutant revealed only a minor defect in the levels of viral RNA replication. To further analyze the consequences of the genomic 3 noncoding region deletion, we examined viral RNA replication in a neuroblastoma cell line, SK-N-SH cells. The minor genomic RNA replication defect in HeLa cells was significantly exacerbated in the SK-N-SH cells, resulting in a decreased capacity for mutant virus growth. Analysis of the nature of the RNA replication deficiency revealed that deleting the poliovirus genomic 3 noncoding region resulted in a positive-strand RNA synthesis defect. The RNA replication deficiency in SK-N-SH cells was not due to a major defect in viral translation or viral protein processing. Neurovirulence of the mutant virus was determined in a transgenic mouse line expressing the human poliovirus receptor. Greater than 1,000 times more mutant virus was required to paralyze 50% of inoculated mice, compared to that with wild-type virus. These data suggest that, together with a cellular factor(s) that is limiting in neuronal cells, the poliovirus 3 noncoding region is involved in positive-strand synthesis during genome replication.Paralytic poliomyelitis is the consequence of the destruction of motor neurons during a poliovirus infection. Only 1 to 2% of individuals infected with poliovirus develop poliomyelitis, suggesting that factors affecting virus tropism and systemic distribution play a significant role in the pathogenic outcome of an infection (7). The cell-and tissue-specific tropism of poliovirus during a systemic infection may be restricted by tissue accessibility (i.e., the blood-brain barrier), cellular attachment and entry, or intracellular interactions between the virus and the cell. In the late 1950s, Holland and colleagues demonstrated that purified poliovirus genomic RNA introduced into nonprimate cells by transfection leads to a single replicative cycle (21). Their data showed that cells which are not normally susceptible to poliovirus infections can be productively infected when receptor-mediated entry of the virus is bypassed, suggesting that the poliovirus receptor (PVR) plays a major role in virus tropism. These conclusions were further supported by the finding that transgenic mice expressing the human PVR (hPVR) are susceptible to poliovirus infection (27,44). Furthermore, the pathology in PVR transgenic mice infected with attenuated or wild-type poliovirus resembles the pathology observed in infected primates, the natural host of poliovirus (22, 43). These observations suggested a strong correlation between PVR-mediated tropism and poliovirus pathology.Although PVR expression may be a major determinant of poliovirus tropism, it is clear that additional factors can influence virus tropism and disease pathology. Assays for virus binding activity in tissue homogenates revealed that binding of virus can occur in tissues where poli...

show abstract

“…The origin and properties of poliovirus strains 452/62 3D, v3/vl-1 and v3/al-25 have been described previously Agol et al, 1984). For the isolation of strains from the CNS of the virusinoculated monkeys, a I0~ suspension of the tissue of the spinal cord lumbar enlargement was prepared in physiological saline and inoculated, in a dose of 0.1 ml, into tubes with replicate monolayer cultures of monkey kidney cells.…”

Section: Methodsmentioning

confidence: 99%

“…M4953, M4973 and M4846, like their parent v3/al-25, failed to synthesize appreciable amounts of viral RNA at this temperature and all should be assigned an RNA-phenotype. In this respect, these strains resembled LSc-gr3, the donor of the 3' half of their genomes (see Agol et al, 1984 …”

Section: Some Phenotypic Properties Of the Isolatesmentioning

confidence: 99%

Neurovirulence of the Intertypic Poliovirus Recombinant v3/a1-25: Characterization of Strains Isolated from the Spinal Cord of Diseased Monkeys and Evaluation of the Contribution of the 3' Half of the Genome

Agol

Drozdov

Frolova

et al. 1985

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYA tsRNA-intertypic recombinant, v3/al-25, which has the 5' and 3' halves of the genome derived from the neurovirulent type 3 poliovirus strain 452/62 3D and the attenuated type 1 poliovirus strain LSc-gr3, respectively, was previously shown to cause severe paralytic poliomyelitis after intracerebral inoculation of monkeys. To ascertain whether the illness was caused by the recombinant itself or by temperature-resistant trRNA ÷ mutants that might have arisen in the inoculated monkeys, five independent virus strains have been isolated from the spinal cord of the diseased animals. While two of these isolates exhibited RNA ÷ and RNA -+ phenotypes, respectively, the other three strains retained the parental RNA-character. Except for the RNA + strain, the RNase T1 oligonucleotide maps of the genomes of all the isolates revealed only a minimal deviation from the parental pattern. These results were interpreted to mean that v3/al-25 is intrinsically neurovirulent despite the presence of a tsRNA-mutation(s) in the 3' half of its genome. Nevertheless, this mutation, or other peculiarities of the 3' half of the recombinant genome, may somewhat alleviate the pathogenicity of the virus. This notion was inferred from the fact that, when used in a relatively small dose (about 103 p.f.u.), v3/al-25 appeared to exhibit a lower level of neurovirulence compared to either the wild-type parent 452/62 3D, or a closely related intertypic recombinant having the genome 3' half derived from a neurovirulent trRNA -+ type 1 poliovirus strain. The problem of genetic determination of poliovirus neurovirulence and attenuation is briefly discussed.

show abstract

Construction and properties of intertypic poliovirus recombinants: First approximation mapping of the major determinants of neurovirulence

Cited by 50 publications

References 12 publications

Analysis of the functional significance of amino acid residues in the putative NTP-binding pattern of the poliovirus 2C protein

Analysis of the functional significance of amino acid residues in the putative NTP-binding pattern of the poliovirus 2C protein

Cell-Dependent Role for the Poliovirus 3′ Noncoding Region in Positive-Strand RNA Synthesis

Neurovirulence of the Intertypic Poliovirus Recombinant v3/a1-25: Characterization of Strains Isolated from the Spinal Cord of Diseased Monkeys and Evaluation of the Contribution of the 3' Half of the Genome

Contact Info

Product

Resources

About