Selection for temperature-sensitive mutations in specific vaccinia virus genes: Isolation and characterization of a virus mutant which encodes a phosphonoacetic acid-resistant, temperature-sensitive DNA polymerase

Sridhar, P.; Condit, Richard C.

doi:10.1016/0042-6822(83)90269-6

Cited by 48 publications

(38 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using this assay, the EC 50 s for CDV and phosphonoacetic acid were determined to be 84 Ϯ 15 M and 985 Ϯ 85 M, respectively. These values were within the range of published values for these compounds (24,37).…”

Section: Cells and Virusessupporting

confidence: 73%

An Orally Bioavailable Antipoxvirus Compound (ST-246) Inhibits Extracellular Virus Formation and Protects Mice from Lethal Orthopoxvirus Challenge

Yang

Pevear²,

Davies³

et al. 2005

J Virol

381

469

View full text Add to dashboard Cite

ST-246 is a low-molecular-weight compound (molecular weight‫؍‬Recent concerns over the use of variola (smallpox) virus as a biological weapon have prompted renewed interest in development of small molecule therapeutics that target variola virus replication. Currently, there is no U.S. Food and Drug Administration-approved drug for the prevention or treatment of smallpox infection. While a number of compounds have been shown to inhibit orthopoxvirus replication in vitro, these compounds often lack potency and/or are associated with significant adverse effects, due to their relative nonspecific mechanisms of virus inhibition (3).The cornerstone of the current national public health response plan to a smallpox bioterrorist attack calls for rapid mass immunization with vaccinia virus. However, concerns about vaccine-related adverse events have compromised implementation of a smallpox immunization program. Individuals with immunodeficiency disorders or certain common skin conditions are unusually susceptible to vaccine-related complications (6, 32). Moreover, the lag period for antibody formation from a vaccine leaves a window of vulnerability. Antiviral therapies can fill this void and provide an excellent complement to vaccination in that they reduce virus titers quickly, regardless of immune status, and lower transmission rates by diminishing the virus reservoir. A small-molecule antiviral drug designed to treat variola virus infection will be a critical component to a smallpox defense strategy.Currently, only cidofovir [CDV; (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine; Vistide], a drug approved for treatment of cytomegalovirus (CMV) retinitis in AIDS pa-* Corresponding author. Mailing address:

show abstract

Section: Cells and Virusessupporting

confidence: 73%

An Orally Bioavailable Antipoxvirus Compound (ST-246) Inhibits Extracellular Virus Formation and Protects Mice from Lethal Orthopoxvirus Challenge

Yang

Pevear²,

Davies³

et al. 2005

J Virol

381

469

View full text Add to dashboard Cite

show abstract

“…The DNA-mutation 1543 mapped in the left-hand one-third of the genome, within an approximately 23 Mdal region. Both these assignments are compatible with the recently reported data of Condit et al (1983) who identified two complementation groups of VV DNA-mutants mapping in the HindIII D and E fragments and with those of Sridhar & Condit (1983) and Jones & Moss (1984) All four 'double pathogenicity" recombinants possessed a single centrally located insert of EMV DNA. For one of these recombinants (R1792-69) it could be deduced that this insert lay within the same 16 Mdal region in which mutations 1792 and 1438 were mapped (Fig.…”

supporting

confidence: 79%

Recombinants between Vaccinia and Ectromelia Viruses Bearing the Specific Pathogenicity Markers of Both Parents

Chernos¹,

Antonova²,

Senkevich³

1985

Journal of General Virology

View full text Add to dashboard Cite

“…Indeed, genetic, genomic, and biochemical analysis has revealed that eight proteins are responsible for vaccinia virus DNA synthesis and maturation. This repertoire includes the catalytic DNA polymerase (E9 (3)(4)(5)(6)(7)(8)(9)(10)(11)), a stoichiometric component of the heterodimeric processivity factor (A20 (12)(13)(14)(15)), a second component of the processivity factor (D4) that also possesses uracil DNA glycosylase (UDG) 2 activity (16 -18), a putative superfamily III helicase with known NTPase and DNA primase activity (D5 (19 -23)), a serine/threonine protein kinase (B1 (24 -26)), an abundant phosphoprotein with essential roles in viral replication, transcription, and morphogenesis (H5 (27)), a single-strand DNA-binding protein (I3 (28,29) 3 ), and a DNA ligase (A50 (30)). A Holliday-junction resolvase (A22 (31)) and a FEN-1 related endonuclease (G5 (32)) have also been shown to be important for formation of monomeric genomes.…”

mentioning

confidence: 99%

Evaluation of the Role of the Vaccinia Virus Uracil DNA Glycosylase and A20 Proteins as Intrinsic Components of the DNA Polymerase Holoenzyme

Boyle

Stanitsa

Greseth

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

The vaccinia virus DNA polymerase is inherently distributive but acquires processivity by associating with a heterodimeric processivity factor comprised of the viral A20 and D4 proteins. D4 is also an enzymatically active uracil DNA glycosylase (UDG). The presence of an active repair protein as an essential component of the polymerase holoenzyme is a unique feature of the replication machinery. We have shown previously that the A20-UDG complex has a stoichiometry of ϳ1:1, and our data suggest that A20 serves as a bridge between polymerase and UDG. Here we show that conserved hydrophobic residues in the N terminus of A20 are important for its binding to UDG. Our data argue against the assembly of D4 into higher order multimers, suggesting that the processivity factor does not form a toroidal ring around the DNA. Instead, we hypothesize that the intrinsic, processive DNA scanning activity of UDG tethers the holoenzyme to the DNA template. The inclusion of UDG as an essential holoenzyme component suggests that replication and base excision repair may be coupled. Here we show that the DNA polymerase can utilize dUTP as a substrate in vitro. Moreover, uracil moieties incorporated into the nascent strand during holoenzyme-mediated DNA synthesis can be excised by the viral UDG present within this holoenzyme, leaving abasic sites. Finally, we show that the polymerase stalls upon encountering an abasic site in the template strand, indicating that, like many replicative polymerases, the poxviral holoenzyme cannot perform translesion synthesis across an abasic site.The faithful and efficient duplication of genomic DNA is one of the most conserved processes across all forms of life. Although this is a necessary and highly regulated process, it seems that each model organism has evolved unique modifications during this process. Members of the poxvirus family, of which variola virus is the most notable member and vaccinia virus is the experimental prototype, are no exception. Poxviruses are unique in that they complete the replication and maturation of their ϳ200-kb double-stranded DNA genome in the cytoplasm of the infected host cell. This autonomy dictates that poxviruses encode many of the proteins necessary for nucleotide precursor synthesis and metabolism as well as the core set of enzymes and DNA binding proteins that act directly at the replication fork (1, 2). Indeed, genetic, genomic, and biochemical analysis has revealed that eight proteins are responsible for vaccinia virus DNA synthesis and maturation. This repertoire includes the catalytic DNA polymerase (E9 (3-11)), a stoichiometric component of the heterodimeric processivity factor (A20 (12-15)), a second component of the processivity factor (D4) that also possesses uracil DNA glycosylase (UDG) 2 activity (16 -18), a putative superfamily III helicase with known NTPase and DNA primase activity (D5 (19 -23)), a serine/threonine protein kinase (B1 (24 -26)), an abundant phosphoprotein with essential roles in viral replication, transcription, and morphogenesis (H5 ...

show abstract

Selection for temperature-sensitive mutations in specific vaccinia virus genes: Isolation and characterization of a virus mutant which encodes a phosphonoacetic acid-resistant, temperature-sensitive DNA polymerase

Cited by 48 publications

References 19 publications

An Orally Bioavailable Antipoxvirus Compound (ST-246) Inhibits Extracellular Virus Formation and Protects Mice from Lethal Orthopoxvirus Challenge

An Orally Bioavailable Antipoxvirus Compound (ST-246) Inhibits Extracellular Virus Formation and Protects Mice from Lethal Orthopoxvirus Challenge

Recombinants between Vaccinia and Ectromelia Viruses Bearing the Specific Pathogenicity Markers of Both Parents

Evaluation of the Role of the Vaccinia Virus Uracil DNA Glycosylase and A20 Proteins as Intrinsic Components of the DNA Polymerase Holoenzyme

Contact Info

Product

Resources

About