Identification of the herpes simplex virus DNA polymerase gene

Purifoy, Dorothy J. M.; Lewis, Robert B.; Powell, Kenneth L.

doi:10.1038/269621a0

Cited by 139 publications

(74 citation statements)

References 11 publications

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“…It is a 1235-amino acid, 136,413-Da protein that is encoded by the UL30 gene (29,(51)(52)(53). The UL30 protein exhibits significant sequence similarity to other viral and cellular DNA polymerases including human DNA polymerase ␣-primase, Saccharomyces cerevisiae DNA polymerase ␦, E. coli DNA polymerase I, and the bacteriophage T4 DNA polymerase (54,55).…”

Section: Hsv-1 Gene Products Essential For Origin-specific Dnamentioning

confidence: 99%

Replication of Herpes Simplex Virus DNA

Lehman

Boehmer

1999

Journal of Biological Chemistry

168

140

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Section: Hsv-1 Gene Products Essential For Origin-specific Dnamentioning

confidence: 99%

Replication of Herpes Simplex Virus DNA

Lehman

Boehmer

1999

Journal of Biological Chemistry

168

140

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“…These were done exactly as described previously (Purifoy & Benyesh-Melnick, 1975;Purifoy et al, 1977).…”

Section: Cells and Virusesmentioning

confidence: 99%

Herpes Simplex Virus Non-structural Proteins. III. Function of the Major DNA-binding Protein

Littler¹,

Purifoy²,

Minson³

et al. 1983

Journal of General Virology

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SUMMARYThe herpes simplex virus type 2 major DNA-binding protein has been functionally characterized using temperature-sensitive mutants in the complementation group 2-2. The mutants were shown to be defective in the DNA-binding protein gene by mapping the mutants to the area of the genome known to code for the protein, and by demonstrating alterations in the major DNA-binding protein induced in mutantinfected cells. The mutants were shown to be defective in the replication of virus DNA. The nature of this defect was examined by studying virus DNA synthesis in vitro and by the examination of virus enzymes. An effect of mutation in the DNA-binding protein was to destabilize both the DNA polymerase and the alkaline exonuclease.

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“…The products of two of these genes, the UL30 gene encoding the 136K viral DNA polymerase (Pol) (Purifoy et al, 1977;Chartrand et al, 1980) and the UL42 gene encoding a 65K double-stranded DNA-binding protein (UL42) , interact within the infected cell to form a heterodimeric complex (Pol/UL42) (Gallo et al, 1988). The use of recombinant baculoviruses to over-express Pol and UL42 as individual subunits has allowed a detailed analysis of the effect of UL42 on Pol activity (Hernandez & Lehman, 1990;Gottlieb et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

The herpes simplex virus type 1 DNA polymerase accessory protein, UL42, contains a functional protease-resistant domain

Hamatake

Bifano

Tenney

et al. 1993

Journal of General Virology

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Herpes simplex virus type 1 encodes its own DNA polymerase (Pol), the product of the UL30 gene, and a polymerase accessory subunit, the product of the UL42 gene, both of which are required for viral DNA replication. Pol and the UL42 protein associate to form a heterodimeric complex (Pol/UL42) which is more active and has a higher processivity than the Pol catalytic subunit alone. The Pol/UL42 complex has been reconstituted by mixing together highly purified Pol and UL42 subunits obtained from recombinant baculovirusinfected cells. We have used polymerase activity on poly(dA):oligo(dT20), a template that the Pol subunit utilizes with low efficiency, to measure the formation of the Pol/UL42 complex. Our data indicate that the association constant for the Pol/UL42 complex is 1 × l0 s M -1. Proteolytic digestions of UL42 were performed to determine whether structural domains of UL42 could be disclosed by differential amino acid accessibilities. The ability of these protease-resistant domains to form a functional complex with Pol was determined by measuring their ability to stimulate Pol activity on poly(dA):oligo(dT20). We have found that trypsin digestion of UL42 in the presence of DNA generates protease-resistant fragments of 28K and 8K which co-elute from a MonoQ column and are able to stimulate Pol activity on poly(dA): oligo(dT20 ). Complex formation of the 28K and 8K tryptic fragments with Pol was also shown by their co-immunoprecipitation with antibody to Pol. It was determined that the 28K fragment of UL42 comprised amino acids 1 to 245 or 1 to 254 of UL42, whereas the 8K fragment started at amino acid 255. Thus, controlled proteolysis of UL42 revealed two closely contiguous structural domains that retained the ability to complex with Pol and stimulate Pol activity.

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Identification of the herpes simplex virus DNA polymerase gene

Cited by 139 publications

References 11 publications

Replication of Herpes Simplex Virus DNA

Replication of Herpes Simplex Virus DNA

Herpes Simplex Virus Non-structural Proteins. III. Function of the Major DNA-binding Protein

The herpes simplex virus type 1 DNA polymerase accessory protein, UL42, contains a functional protease-resistant domain

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