Genetic evidence for involvement of vaccinia virus DNA-dependent ATPase I in intermediate and late gene expression

Künzi, Myriam S.; Traktman, Paula

doi:10.1128/jvi.63.9.3999-4010.1989

Cited by 16 publications

(13 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Quantitation of viral DNA replication. The dot blot filter hybridization technique was used to quantitate the accumulation of viral DNA sequences after infection, as described previously (25,35). In temperature shift experiments, confluent 35-mm-diameter dishes of mouse L cells were infected with ts2 and ts25 at a multiplicity of infection (MOI) of 15 PFU per cell.…”

Section: Methodsmentioning

confidence: 99%

Vaccinia virus B1 kinase: phenotypic analysis of temperature-sensitive mutants and enzymatic characterization of recombinant proteins

Rempel

Traktman

1992

J Virol

Self Cite

109

View full text Add to dashboard Cite

The vaccinia virus Bi gene encodes a 34-kDa protein with homology to protein kinases. In L cells infected nonpermissively with mutants containing lesions in the Bi gene (ts2 and ts25), the infectious cycle arrests prior to DNA replication. In this report, we demonstrate that DNA synthesis ceases when cultures infected with these mutants at 32°C are shifted to the nonpermissive temperature (39.5°C) in the midst of DNA replication. We also show that Bi protein is synthesized transiently during the early phase of infection, even when the progression to later stages of gene expression is prevented. Although wild-type (wt) Bl is stable, the ts Bi proteins are markedly labile in both L and BSC40 cells at both permissive and nonpermissive temperatures. These results suggest that the ts phenotype of the mutants is complex and may in part reflect a temperature-dependent requirement for kinase activity, an induction of temperature sensitivity in Bi substrates under nonpermissive

show abstract

Section: Methodsmentioning

confidence: 99%

Vaccinia virus B1 kinase: phenotypic analysis of temperature-sensitive mutants and enzymatic characterization of recombinant proteins

Rempel

Traktman

1992

J Virol

Self Cite

109

View full text Add to dashboard Cite

show abstract

“…At the indicated times postinfection, cultures were placed on ice, rinsed with PBS, and lysed with guanidinium thiocyanate (4 M guanidinium thiocyanate, 25 mM sodium citrate [pH 7.0], 0.5% lauroylsarcosine, 0.1 M P-mercaptoethanol). Lysates were passed four times through a 23-gauge needle, and total RNA was isolated by ultracentrifugation through cesium chloride (5.7 M CsCl, 0.1 M EDTA [pH 7.0]) as described previously (13,21). Purified RNA was stored in distilled H20 at -80°C.…”

Section: Methodsmentioning

confidence: 99%

“…were incubated with a molar excess of DNA probe (25 ng at 600 cpm/ng) under standard denaturing conditions (21,45). Samples were then incubated at 14WC (12 to 15 h) to allow for hybridization, followed by digestion with S1 endonuclease (800 U/ml in 0.28 M NaCI-0.05 M sodium acetate [pH 4.6]-0.5 mM ZnSO4-20 ,ug of sonicated salmon sperm DNA per ml) at 14WC for 1 h to remove nonhybrids.…”

Section: Methodsmentioning

confidence: 99%

Transient expression of the vaccinia virus DNA polymerase is an intrinsic feature of the early phase of infection and is unlinked to DNA replication and late gene expression

McDonald

Crozel-Goudot

Traktman

1992

J Virol

Self Cite

View full text Add to dashboard Cite

We have studied the expression pattern of the vaccinia virus DNA polymerase during the viral replicative cycle. To monitor polymerase synthesis, a polyclonal antiserum was raised against a TrpE-DNA polymerase fusion protein. Immunoprecipitation and Sl analyses revealed that polymerase synthesis and mRNA levels peak by 2 to 3.5 h postinfection during wild-type infections and then decline, becoming barely detectable by 5 to 6.5 h postinfection. Blocking viral DNA replication by performing infections with temperature-sensitive DNA-mutants at the nonpermissive temperature or by performing wild-type infections in the presence of cytosine ,B-D-arabinofuranoside had no effect on polymerase expression. These results indicate that the transient expression of the DNA polymerase is regulated independently of intermediate and late viral gene expression. Cycloheximide, which inhibits protein synthesis and prevents secondary uncoating, caused prolonged and elevated levels of polymerase transcription. Early viral proteins and uncoating, rather than exhaustion of the encapsidated transcription machinery, are presumed to mediate the cessation of polymerase transcription. In the presence of aphidicolin, the polymerase transcripts were maintained at maximal levels rather than exhibiting their normal decline. This inhibition of RNA decay was seen even in infections performed with isolates encoding aphidicolin-resistant DNA polymerases, suggesting that aphidicolin may interfere directly with the process of RNA degradation. Under these conditions, polymerase synthesis remained transient and was not prolonged, despite the continuing presence of available mRNA. These observations suggest that early mRNAs may experience a loss in translation efficiency as infection progresses.

show abstract

“…It is worth noting that subsequent to submission of our initial report (Henikoff, 1993), SNF2_YEAST was revealed to be a DNA-stimulated ATPase without detectable helicase activity (Laurent et al, 1993). Lack of helicase activity is a feature of well-studied members of the poxvirus DNA-dependent ATPases (e. g. Kunzi and Traktman, 1989).…”

Section: Classification Of Suspected Helicasesmentioning

confidence: 96%

Protein Family Classification Based on Searching a Database of Blocks

1994

View full text Add to dashboard Cite

Genetic evidence for involvement of vaccinia virus DNA-dependent ATPase I in intermediate and late gene expression

Cited by 16 publications

References 46 publications

Vaccinia virus B1 kinase: phenotypic analysis of temperature-sensitive mutants and enzymatic characterization of recombinant proteins

Vaccinia virus B1 kinase: phenotypic analysis of temperature-sensitive mutants and enzymatic characterization of recombinant proteins

Transient expression of the vaccinia virus DNA polymerase is an intrinsic feature of the early phase of infection and is unlinked to DNA replication and late gene expression

Protein Family Classification Based on Searching a Database of Blocks

Contact Info

Product

Resources

About