Properties of Herpes simplex virus type 1 and type 2 DNA polymerase

Ostrander, Michael; Cheng, Yung‐Chi

doi:10.1016/0005-2787(80)90234-8

Cited by 118 publications

(74 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This agrees closely with values reported by Furman et al (1979) when dGTP was used as substrate for DNA polymerase from four different strains of HSV-1, although it is somewhat greater than the value reported recently by Ostrander & Cheng (1980). When dTTP was the limiting substrate, however, we observed biphasic kinetics showing substrate activation occurring at the higher substrate concentrations used (Fig.…”

Section: Discussionsupporting

confidence: 93%

“…6), although the K m value increased from 0.5 to 5 /~M. Ostrander & Cheng (1980) reported only a single K m value of 0.14/~M for dTTP for the HSV-l-induced DNA polymerase. The different kinetic behaviour observed by these workers, compared with our results, possibly reflects differences in the purity of the polymerases and in the conditions of the assays.…”

Section: Discussionmentioning

confidence: 92%

See 1 more Smart Citation

Aspects of Thymidine Metabolism and Function in Cultured Mammalian Cells Infected with Herpes Simplex Virus Type 1

Baybutt¹,

Murray²,

Pearson³

1982

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYWe have investigated aspects of thymidine metabolism and function in cultured mammalian cells infected with herpes simplex virus under conditions in which virus DNA synthesis was either permitted, or was prevented by inhibiting the virus-induced DNA polymerase with phosphonoacetate. In 30 min pulse-labelling experiments the rates of [3H]thymidine transport into cells and its subsequent phosphorylation both reached maximum values about 6 h after infection. During the next 17 h these rates remained relatively constant in the absence of phosphonoacetate but declined to about 50% of the 6 h value in the presence of this inhibitor. When the radioactive thymidine was present continuously throughout the infection the accumulated intracellular acid-soluble radioactivity reached maximum values at about 9 h. In the absence of phosphonoacetate this declined thereafter, but it remained relatively constant when virus DNA synthesis was prevented. This suggests some established equilibrium between the continuing entry of thymidine into cells and its subsequent removal by excretion. Excretion of thymidine-derived radioactivity was initially established by determining the fate of isotopically labelled host cell DNA. During 24 h of infection about 50% of the host cell DNA was rendered acid-soluble and of this 60 to 70% was excreted from the cells. High pressure liquid chromatographic analysis of the intracellular acid-soluble radioactivity showed only phosphorylated thymidine derivatives, whereas the excreted radioactivity was exclusively in thymidine. Excretion of intracellular radioactive molecules derived directly from [3H]thymidine in the medium was also observed, but only when virus DNA synthesis was inhibited with phosphonoacetate.Simple kinetic studies with the purified virus-induced DNA polymerase showed that a straight line double-reciprocal plot was obtained when dGTP was used as the limiting substrate (Km = 0.8 aM, Vmax = 330 nmol dGTP incorporated/10 min) but that a biphasic plot was obtained when dTTP was limiting (Kml ----0.5 aM, Vmaxl = 208 nmol dTTP incorporated/10 min; Kin2 = 5 aM, Vmax2 = 370 nmol dTTP incorporated/10 rain).

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 92%

Aspects of Thymidine Metabolism and Function in Cultured Mammalian Cells Infected with Herpes Simplex Virus Type 1

Baybutt¹,

Murray²,

Pearson³

1982

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…6). The identical kinotics of heat denaturation were reported earlier for polymerase and exonuclease activities of phage 029 DNA polymerase ( 25 ), herpes virus DIA polymerase ( 10,12 ), vaccinia virus DNA polymerase ( 13 ) and alian DNA polymerase 6S ( 15 ). The aphidicolin-sensitivity of 3'*5' exonuclease also indicates that the exonuclease is associated with BINPV Pol.…”

Section: Materials and Methods Chemicals And Enzymessupporting

confidence: 78%

“…No data were published on exonucleases associated with DNA polymerases of baculoviruses. However, the intrinsic 3'*5' exonucleases were found to be associated with DNA polymerases of herpes viruses (8)(9)(10)(11)(12), vacc.inia virus ( 13 ) and adenoviruses ( 14 ). In this respect, DNA polymerases of animal DNA viruses resemble prokaryotic enzymes.…”

Section: Nucleic Acids Researchmentioning

confidence: 99%

Characterization of 3'→5' exonuclease associated with DNA polymerase of silkworm nuclear polyhedrosis virus

Mikhailov

Marlyev²,

Ataeva³

et al. 1986

Nucl Acids Res

View full text Add to dashboard Cite

“…7c), a much more pronounced difference in PAA-sensitivity of the wild-type and HVS (pr) virus polymerases was observed; the 50~ inhibitory concentrations of PAA for these activities in extracts of cells infected with sensitive and resistant virus were 2 and 25 txg/ml, respectively. The mechanism of these salt-dependent effects on the apparent PAA-resistance of the HSV-and HVS-induced activities is uncertain, but similar phenomena have been noted by others with both crude and partially purified HSV-specified enzyme activities (Ostrander & Cheng, 1980).…”

Section: Induction Of a Novel Aphidicolin-resistant Paa-sensitive Dnsupporting

confidence: 60%

Evidence for a Herpesvirus Saimiri-specified DNA Polymerase Activity which is Aphidicolin-resistant and Phosphonoacetate-sensitive

O’Hare¹,

Honess²

1983

Journal of General Virology

View full text Add to dashboard Cite

SUMMARYPhosphonoacetic acid (PAA) effectively inhibited herpesvirus saimiri (HVS) replication and the onset of virus DNA synthesis. A PAA-resistant (pr) mutant of HVS was isolated which plaqued efficiently in the presence of concentrations of PAA sufficient to reduce the growth of wild-type virus to < 0-02~ of control values. In contrast, virus growth and DNA synthesis in cells infected with unselected strains of herpesvirus saimiri was highly resistant to concentrations of aphidicolin, an inhibitor of a-type polymerases, which completely inhibited the growth and DNA replication of uninfected cells. An increased level of DNA polymerase activity was induced in cells infected with herpesvirus saimiri. This HVS-induced DNA polymerase was more sensitive to PAA but more resistant to aphidicolin in vitro than the uninfected cell activity and could also be distinguished on the basis of its response to ionic strength (40 to 50 raM-ammonium sulphate for optimal activity versus 20 mM for the uninfected cell activity). Under suitable in vitro assay conditions, an increase in the PAA-resistance of the DNA polymerase induced by the HVS(P0 mutant was demonstrated. Comparison of the effects of aphidicolin and PAA demonstrated that the former was a much more effective and rapid inhibitor of susceptible cell DNA synthesis in vivo. Taken together, these results demonstrate novel properties of a DNA polymerase activity in cells infected with herpesvirus saimiri and suggest that aphidicolin should provide a useful reagent to analyse the functions of this enzyme in productive and non-productive infections with the virus.

show abstract

Properties of Herpes simplex virus type 1 and type 2 DNA polymerase

Cited by 118 publications

References 29 publications

Aspects of Thymidine Metabolism and Function in Cultured Mammalian Cells Infected with Herpes Simplex Virus Type 1

Aspects of Thymidine Metabolism and Function in Cultured Mammalian Cells Infected with Herpes Simplex Virus Type 1

Characterization of 3'→5' exonuclease associated with DNA polymerase of silkworm nuclear polyhedrosis virus

Evidence for a Herpesvirus Saimiri-specified DNA Polymerase Activity which is Aphidicolin-resistant and Phosphonoacetate-sensitive

Contact Info

Product

Resources

About