Mapping of the varicella zoster virus deoxypyrimidine kinase gene and preliminary identification of its transcript

Sawyer, Mark H.; Ostrove, Jeffrey M.; Felser, James M.; Straus, Stephen E.

doi:10.1016/0042-6822(86)90081-4

Cited by 33 publications

(17 citation statements)

References 32 publications

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“…As previously noted, the VZV dPK exhibits greater deoxycytidine kinase activity than thymidine kinase activity (Sawyer et al, 1986). Strain KB3 was also shown to exhibit acyclovir-phosphorylating activity that was less than 2% of that of its wt parent, VZV Ellen (data not shown).…”

Section: Resultssupporting

confidence: 64%

“…1. The VZV dPK transcript has been characterized and is 1.8 kb in size (Sawyer et al, 1986;Davison & Scott, 1986). Northern blot analyses, in which infected cellular RNAs were probed with the 700 bp AvaI VZV DNA subfragment that lies completely within the dPK gene, demonstrated that wt and acyclovir-resistant mutants all produced the 1.8 kb 1-5 0-9 VZV 101 0.54 1.7 * Activity is expressed relative to that of uninfected cells.…”

Section: Resultsmentioning

confidence: 99%

“…The precise mapping of the VZV dPK gene (Sawyer et al, 1986;Davison & Scott, 1986) allowed us to isolate, clone and sequence the gene from both drug-sensitive and drug-resistant VZV strains and to identify the mutations that render them acyclovir-resistant. The VZV dPK open reading frame consists of 1023 bp and encodes a protein of 341 amino acids (Davison & Scott, 1986).…”

Section: Discussionmentioning

confidence: 99%

“…Five pellets were prepared from each flask and stored at -70 °C until assayed. The dPK activity in cell lysates was determined by measuring the in vitro phosphorylation of [3H]thymidine and [3H]deoxycytidine as previously described (Sawyer et al, 1986). The enzyme activity was normalized for the amount of protein in each extract.…”

Section: Methodsmentioning

confidence: 99%

“…The VZV dPK gene has been mapped to the middle of the unique long (UL) segment of the VZV genome (map units 0.50 to 0.52) and shown to encode a 1.8 kb transcript (Sawyer et al, 1986: Davison & Scott, 1986. The herpes simplex virus (HSV) thymidine kinase (TK) shares approx.…”

Section: Introductionmentioning

confidence: 99%

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Molecular Analysis of the Pyrimidine Deoxyribonucleoside Kinase Gene of Wild-type and Acyclovir-resistant Strains of Varicella-Zoster Virus

Sawyer

Inchauspé

Biron

et al. 1988

Journal of General Virology

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SUMMARYThe pyrimidine deoxyribonucleoside kinase (dPK) genes from five wild-type and four acyclovir-resistant varicella-zoster virus (VZV) strains were studied. One of the acyclovir-resistant strains was isolated from a patient receiving chronic acyclovir therapy. Acyclovir-resistant strains expressed the 1.8 kb VZV dPK transcript but lacked dPK activity. To determine the basis for the lack of enzyme activity the dPK gene from each strain was cloned and its DNA sequence determined. The VZV dPK gene was found to be highly conserved among strains, with greater than 99 % nucleotide and amino acid homology. Each acyclovir-resistant VZV strain differed from its wildtype parent in only a single amino acid. The dPK genes from the acyclovir-resistant strains contained either point mutations near the putative thymidine-binding site of the enzyme or ones that resulted in the premature termination of protein synthesis. Single point mutations were sufficient to render these strains dPK-negative and highly resistant to acyclovir. The molecular basis for acyclovir resistance at the dPK locus of VZV is similar to that previously noted to render herpes simplex viruses resistant to acyclovir.

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Section: Resultssupporting

confidence: 64%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular Analysis of the Pyrimidine Deoxyribonucleoside Kinase Gene of Wild-type and Acyclovir-resistant Strains of Varicella-Zoster Virus

Sawyer

Inchauspé

Biron

et al. 1988

Journal of General Virology

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Analyses of the transcriptional pattern of glycoproteins E and I of varicella-zoster virus and evidence for a monocistronic transcription

Rahaus

Wolff

2003

J. Med. Virol.

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Glycoproteins I and E of the Varicella-zoster virus, encoded by the neighbouring open reading frames 67 and 68, are transcribed into several transcript species that differ in size. From gI, three transcripts of 1.65, 2.7, and 3.6 kb are known, and from gE, two transcripts of 2.15 and 3.6 kb in size are known. Here, we demonstrate that these various transcript species appear in different amounts at different times post infection. At 12 hr post infection, the transcripts of 1.65 (gI) and 2.15 (gE) were clearly detectable, whereas the other transcripts appeared later on. RT-PCR studies using a set of different primers provided clear evidence that gI and gE are transcribed both, mono- and bicistronically, with predominance on the respective monocistronic transcript. Additional evidence for monocistronic transcription was found in the fact that both glycoproteins contain their own transcriptional start sites. Both promoter regions have their own basal transcription activity and include active TATA-boxes that were recognized by the TATA-box binding protein.

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Herpesviridae: Varicella-Zoster Virus

Takahashi¹

1988

Laboratory Diagnosis of Infectious Diseases Principles and Practice

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Mapping of the varicella zoster virus deoxypyrimidine kinase gene and preliminary identification of its transcript

Cited by 33 publications

References 32 publications

Molecular Analysis of the Pyrimidine Deoxyribonucleoside Kinase Gene of Wild-type and Acyclovir-resistant Strains of Varicella-Zoster Virus

Molecular Analysis of the Pyrimidine Deoxyribonucleoside Kinase Gene of Wild-type and Acyclovir-resistant Strains of Varicella-Zoster Virus

Analyses of the transcriptional pattern of glycoproteins E and I of varicella-zoster virus and evidence for a monocistronic transcription

Herpesviridae: Varicella-Zoster Virus

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