Inhibition of avian myeloblastosis virus deoxyribonucleic acid polymerase by synthetic polynucleotides

Erickson, Robert J.; Grosch, Josephine C.

doi:10.1021/bi00706a032

Cited by 35 publications

(11 citation statements)

References 19 publications

(13 reference statements)

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“…If this system is to be analogous to that reported for the oncornaviruses (8,16,19,21), then (dUfl)n should inhibit the activity ofthe virus-encapsulated nucleic acid polymerase. Purified B particles were solubilized by the high salt method of Emerson and Wagner (6) and assayed for RNA polymerase activity using the endogenous viral genome as template.…”

Section: Resultsmentioning

confidence: 81%

“…This area of investigation gained additional impetus with the simultaneous development of a number of nuclease-resistant forms of synthetic polynucleotides including (1-vinyluracil), and (9-vinyladenine)n (16), (2'-O-methyladenylic acid), (21), and (dUfl)n (8), all of which have been found to inhibit oncornavirus DNA polymerase activity.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, a more direct mechanism of antiviral activity has been attributed to this class of polymers by the demonstration that certain single-stranded polynucleotides can block the activity of virus-associated nucleic acid polymerases (8,16,17,22). The majority of these data was obtained in studies utilizing the ribonuleic acid (RNA)-dependent deoxyribonucleic acid (DNA) polymerase ofthe oncornaviruses and indicated that the inhibitory polynucleotides blocked the association of enzyme and active nucleic acid template (7).…”

mentioning

confidence: 99%

“…[(dUfl)J], a potent inhibitor of the avian myeloblastosis virus RNA-dependent DNA polymerase (8). This system appeared to be amenable to our studies since VS virus carries a viral-specific, RNA-dependent RNA polymerase, viral replication is relatively independent of host cell DNA and RNA synthesis, and (dUfl), was found to be nontoxic to the host cells.…”

mentioning

confidence: 99%

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Inhibition of Vesicular Stomatitis Virus Replication by Poly(2′-Fluoro-2′-Deoxyuridylic Acid)

Grosch

Erickson

1975

Antimicrob Agents Chemother

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Poly(2'-fluoro-2'-deoxyuridylic acid) is known to be an effective inhibitor ofthe deoxyribonucleic acid polymerase found within the oncornaviruses. This synthetic polynucleotide was found to inhibit the replication of vesicular stomatitis virus in mouse L cells. The polymer was shown to be capable of inhibiting the viral ribonucleic acid (RNA)-dependent RNA polymerase, and it is proposed that this is the mechanism of antiviral activity. The following observations support this viewpoint: (i) the polymer is most active when added after virus adsorption; (ii) the antiviral activity is not species specific; and (iii) the polynucleotide is nontoxic to the host cell. Conventional methodologies designed to increase nucleic acid uptake by cultured cells do not show an increase in antiviral potency.The antiviral properties of synthetic polynucleotides have, in the past, been associated with either their ability to induce interferon (23) or to enhance the immune response against viral antigens (3). Recently, a more direct mechanism of antiviral activity has been attributed to this class of polymers by the demonstration that certain single-stranded polynucleotides can block the activity of virus-associated nucleic acid polymerases (8,16,17,22). The majority of these data was obtained in studies utilizing the ribonuleic acid (RNA)-dependent deoxyribonucleic acid (DNA) polymerase ofthe oncornaviruses and indicated that the inhibitory polynucleotides blocked the association of enzyme and active nucleic acid template (7). Although the extension of these enzyme analyses to cell culture (16,19,21) and in vivo virus inhibition studies (20) did demonstrate significant antiviral activity, the actual mechanism of inhibition remains unclear. The replication of the oncornaviruses is a complex process that is strongly dependent on host cell metabolism (18), and mechanistic studies involving these viruses may remain difficult and equivocal.Efforts to find a less complex virus-host cell system in which to study the direct inhibitory -activity of the single-stranded polymers demonstrated that the replication ofvesicular stomatitis (VS) virus in mouse L-cell culture was sensitive to poly(2'-fluoro-2'-deoxyuridylic acid)[(dUfl)J], a potent inhibitor of the avian myeloblastosis virus RNA-dependent DNA polymerase (8). This system appeared to be amenable to our studies since VS virus carries a viral-specific, RNA-dependent RNA polymerase, viral replication is relatively independent of host cell DNA and RNA synthesis, and (dUfl), was found to be nontoxic to the host cells. The present study demonstrates that the VS virus RNA polymerase is sensitive to (dUfl)!,, inhibition and the polynucleotide is active when administered postadsorption and presents data which suggest that the inhibition is not mediated by interferon.(A preliminary report on these investigations was presented at the Fourteenth Interscience

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Section: Resultsmentioning

confidence: 81%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Inhibition of Vesicular Stomatitis Virus Replication by Poly(2′-Fluoro-2′-Deoxyuridylic Acid)

Grosch

Erickson

1975

Antimicrob Agents Chemother

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“…Whereas (A)n, (C)n, and (I)n, when appropriately primed, are efficient templates for RNA-dependent DNA polymerase (2), (U)n is a potent competitive inhibitor when (C)n-(dG)i2i8 is treated as the variable substrate. The inhibitory properties of (U)n are markedly enhanced by modification at the 2' position (3,4). It also has been reported that the replication of Moloney leukemia virus in cell cultures of NIH Swiss strain embryo cells is inhibited by (A)n (5).…”

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confidence: 97%

Reactivity of reverse transcriptase toward (s4U,U)n copolymers and spin-labeled nucleic acid lattices.

Warwick¹,

Hakam²,

Bobst³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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Spin-labeled copolymers of 4-thiouridine and uridine [(ts4UU)] that contain various amounts of spin label (e) were synthesized by either (i) chemical alkylation of the 4-thiouridine-uridine copolymers (s4U,U). prepared by copolymerizing 4-thiouridine 5'-diphosphate (s4UDP) and UDP or (ii) copolymerization of spin-labeled S4UDP with UDP using polynucleotide phosphorylase. The effect of (s4U,U). and (tS4UU). on avian myeloblastosis virus (AMV) RNA-dependent DNA polymerase (RNA-dependent DNA nucleotidyltransferase, EC 2.7.7.7; reverse transcriptase) was studied to determine whether the presence of potentially reactive thiol groups or spin labels enhances the inhibitory properties of the copolymers as compared to (U)3. Inhibition by (s4U,U) gradually increases as the percentage of thiolation increases. Enhanced inhibition by (s4U,U). appears to be due to the interaction of the thiol groups of (s4U,U). with the thiol group(s) of the polymerase, because inhibition by (s4U,U). (8% thiolated) in the presence of dithiotreitol resembles that by (U)L. In contrast, inhibition by (ts4UU). containing 3% spin label resembles that by (U)L; however, increasing the spin label to 6% or 12% results in enhanced inhibition by (4s4U,U). as compared to that by (U)3, and dithiothreitol has no effect on enhanced inhibition by (t UU)3.These results suggest that the mechanism of inhibition observed with (ts4U,U). with a es4U:U ratio > 1:33 differs from the mechanism for (s4U,U). and involves complex formation between the spin label and the essential Zn2+ of RNA-dependent DNA polymerase.

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Effect of polyamines on the uptake of poly (2′‐fluoro‐2′‐deoxyuridylic acid) by a mammalian cell line

Janik

Sommer

1976

J Supramol Struct

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VERO cells can take up poly(dUfl)1 from the medium. The uptake involves surface adsorption and, most probably, intracellular penetration. Part of the poly(dUfl) is hydrolyzed during incubation with the cells but the hydrolysis products are not incorporated into de novo synthesized nucleic acids. The uptake is reduced by serum and stimulated by polycationic ionenes. The magnitude of stimulation depends on the structure of the ionene and the treatment regimen.

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Inhibition of avian myeloblastosis virus deoxyribonucleic acid polymerase by synthetic polynucleotides

Cited by 35 publications

References 19 publications

Inhibition of Vesicular Stomatitis Virus Replication by Poly(2′-Fluoro-2′-Deoxyuridylic Acid)

Inhibition of Vesicular Stomatitis Virus Replication by Poly(2′-Fluoro-2′-Deoxyuridylic Acid)

Reactivity of reverse transcriptase toward (s4U,U)n copolymers and spin-labeled nucleic acid lattices.

Effect of polyamines on the uptake of poly (2′‐fluoro‐2′‐deoxyuridylic acid) by a mammalian cell line

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