Identification of strains of herpes simplex virus by comparison of the density of their DNA using the preparative ultracentrifuge

Halliburton, I. W.; Hill, Elizabeth A.; Russell, G. J.

doi:10.1007/bf01318148

Cited by 8 publications

(4 citation statements)

References 34 publications

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“…Our complete sequence for HSV-1 DNA contains 152260 residues in each strand, and has a base composition of 68.3~ G+C. This is slightly higher than the value of 67 ~, based on buoyant density centrifugation, which has been accepted in recent years (Kieffet al, 1971 ;Halliburton et al, 1975). As indicated in Fig.…”

Section: Overall View Of the Hsv-1 Genome Sequencementioning

confidence: 80%

The Complete DNA Sequence of the Long Unique Region in the Genome of Herpes Simplex Virus Type 1

McGeoch

Dalrymple

Davison

et al. 1988

Journal of General Virology

1,574

1,230

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We have determined the DNA sequence of the long unique region (UL) in the genome of herpes simplex virus type 1 (HSV-1) strain 17. The UL sequence contained 107943 residues and had a base composition of 66-9~ G+C. Together with our previous work, this completes the sequence of HSV-1 DNA, giving a total genome length of 152260 residues of base composition 68.3~ G+C. Genes in the UL region were located by the use of published mapping analyses, transcript structures and sequence data, and by examination of DNA sequence characteristics. Fifty-six genes were identified, accounting for most of the sequence. Some 28 of these are at present of unknown function. The gene layout for UL was found to be very similar to that for the corresponding part of the genome of varicella-zoster virus, the only other completely sequenced alphaherpesvirus, and the amino acid sequences of equivalent proteins showed a range of similarities. In the whole genome of HSV-1 we now recognize 72 genes which encode 70 distinct proteins. INTRODUCTION In the last decade, the study of animal viruses has been revolutionized by the application of nucleic acid sequencing techniques to viral genomes. Many smaller virus genomes have been completely sequenced, and the sequences interpreted to give high resolution views of the genetic organization and the nature of the encoded proteins, while comparisons of sequences have enhanced our understanding of relationships between viruses. For larger virus genomes, total determination of nucleotide sequence remains a formidable undertaking, and only two complete sequences of virus genomes larger than 105 residues have been published. These are for the gammaherpesvirus Epstein-Barr virus (EBV) of 172282 residues (Baer et al., 1984) and the alphaherpesvirus varicella-zoster virus (VZV) of 124884 residues (Davison & Scott, 1986a). In this paper we report a third complete herpesvirus genome sequence, that of herpes simplex virus type 1 (HSV-1), which comprises 152260 residues. The molecular biology and genetics of HSV types 1 and 2 have been widely investigated such that overall they are the most extensively characterized of the family Herpesviridae. A decade ago, studies on the structure of HSV DNA showed it to be a linear molecule which could be viewed as consisting of two covalently linked segments, designated long (L) and short (S). Each segment contains a unique sequence flanked by a pair of inverted repeat sequences, as shown in Fig. 1. The long repeat (RL) and short repeat (Rs) sequences are distinct. The molecule also

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Section: Overall View Of the Hsv-1 Genome Sequencementioning

confidence: 80%

The Complete DNA Sequence of the Long Unique Region in the Genome of Herpes Simplex Virus Type 1

McGeoch

Dalrymple

Davison

et al. 1988

Journal of General Virology

1,574

1,230

View full text Add to dashboard Cite

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“…HSV-I DNA was isolated by phenol/SDS extraction of pelleted cell released virus (Wilkie, I973) after purification of the virus on gradients of dextran T-Io (Spear & Roizman, 1972). All DNA preparations used, when analysed by isopycnic banding in CsC1 density gradients, contained a single detectable DNA component with a density characteristic of HSV-I DNA (Halliburton, Hill & Russell, 1975).…”

Section: Cells and Virusmentioning

confidence: 99%

A Partial Denaturation Map of Herpes Simplex Virus Type 1 DNA: Evidence for Inversions of the Unique DNA Regions

Delius

Clements

1976

Journal of General Virology

151

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SUMMARYPartial denaturation maps of 30 HSV-I DNA molecules have been obtained using a procedure designed to avoid possible hydrolysis of the DNA at alkalilabile bonds. From the denaturation pattern of the long unique DNA region these molecules were divided into two groups comprised of 16 and 14 molecules. Histogram plots relating the percentage denaturation to position on the DNA for these two groups were aligned in a manner appropriate to the HSV-I genome model. It was apparent that these groups had the orientation of the long region inverted with respect to each other. Similarly, from the denaturation maps of the short unique region, the molecules were divided into two groups each comprising 15 molecules. Alignment of the histogram plots of these groups indicated that the orientation of the short region was inverted in one group relative to the other. These partial denaturation data confirm the presence of four HSV-I genome arrangements resulting from the possible combinations of inversions of the two unique DNA regions.

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“…In particular we report in this paper evidence that ara-T: (i) is phosphorylated by extracts of HSV-infected BHK cells but not by those of uninfected BHK cells; (ii) 25 ,ug of BrdU per ml.…”

mentioning

confidence: 56%

“…CsCl density gradient centrifugation was used to separate viral from cellular DNA by minor modifications of the procedure of Halliburton et al (25). Replicate confluent monolayers in 24-well plates were infected with 50 ,ul of HSV-1 (MOI = 1).…”

mentioning

confidence: 99%

Antiviral Activity of Arabinosylthymine in Herpesviral Replication: Mechanism of Action In Vivo and In Vitro

Aswell¹,

Allen²,

Jamieson³

et al. 1977

Antimicrob Agents Chemother

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1-p-D-Arabinofuranosylthymine (ara-T), a metabolite of the sponge Tethya crypta, has shown selective activity against herpes simplex virus (HSV) replication (G. A. Gentry and J. F. Aswell, Virology 65:294-296, 1975 responsible for the selective antiviral activity of ara-T. This conclusion was further supported by experiments that showed that the replication ofa variety of dPyK-mutants of HSV-1 and HSV-2 were not affected by ara-T and that ara-T inhibited the phosphorylation of deoxycytidine and deoxythymidine by HSV-1 dPyK, but not by host deoxycytidine and deoxythymidine kinases, respectively. Ara-T also selectively inhibited the replication ofequine herpesvirus type 1 (EHV-1) in vitro and was effective against EHV-1 infection in vivo in hamsters. Further, EHV-1 was inhibited by ara-T and by bromodeoxyuridine in LM cells lacking a cytosol thymidine kinase, suggesting that EHV-1 induces a dPyK. Finally, spectrophotometric assay for thymine suggested that ara-T is not a substrate for nucleoside phosphorylase of hamster liver, and a microbiological assay indicated that substantial amounts of ara-T were excreted in the urine of uninfected hamsters that had received a single injection of 5 mg of ara-T, the amount given in each injection in the in vivo experiments with EHV-1.1-p-D-Arabinofuranosylthymine (ara-T; spongothymidine) was first isolated, rather by coincidence, by Bergmann and Feeney (5) in a study of the sterol content (4) of Tethya crypta, a sponge first described as growing almost buried in the ocean floor off the Bahamas (16). Subsequently, ara-T was found to have little effect on the multiplication of tumor cells in culture (11,15) but was later shown to be somewhat inhibitory to herpesvirus replication in vivo (17,18,53,54,56). The demonstration (26) that herpes simplex virus (HSV)-infected cells contain a pyrimidine deoxyribonucleoside kinase (dPyK) with a broad substrate specificity (29,30) suggested that ara-T might be phospho- showing that ara-T at 2 x 10-M completely blocked the replication of HSV type 1 (HSV-1) and HSV type 2 (HSV-2) in BHK cells but had no measurable effect on the multiplication of uninfected BHK cells subcultured at low cell density and followed for several generations (23). It has subsequently been shown that several other deoythymidine (dT) analogs have similar selectivity against HSV, including 5'-amino-2',5'-dideoxy-5-iodouridine, 5-ethyldeoxyuridine, 5-propyldeoxyuridine, 5-allyldeoxyuridine (8, 10); 5-bromodeoxycytidine, 5-iododeoxycytidine (13,20, 24,31); and 5-methoxymethyldeoxyuridine (2,3). This suggests that the HSV-specified dPyK may be a useful tool in

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Identification of strains of herpes simplex virus by comparison of the density of their DNA using the preparative ultracentrifuge

Cited by 8 publications

References 34 publications

The Complete DNA Sequence of the Long Unique Region in the Genome of Herpes Simplex Virus Type 1

The Complete DNA Sequence of the Long Unique Region in the Genome of Herpes Simplex Virus Type 1

A Partial Denaturation Map of Herpes Simplex Virus Type 1 DNA: Evidence for Inversions of the Unique DNA Regions

Antiviral Activity of Arabinosylthymine in Herpesviral Replication: Mechanism of Action In Vivo and In Vitro

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