The Vaccinia Virus D5 Protein, Which is Required for DNA Replication, is a Nucleic Acid-independent Nucleoside Triphosphatase

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1995 Sep 1

PMID 7636979

Citations 48

Authors

E Evans

N Klemperer

R Ghosh

P Traktman

Affiliations

Soon will be listed here.

Abstract

The vaccinia virus D5 gene encodes a 90-kDa protein that is transiently expressed at early times after infection. Temperature-sensitive mutants with lesions in the D5 gene exhibit a fast-stop DNA- phenotype and are also impaired in homologous recombination. Here we report the overexpression of the D5 protein within the context of a vaccinia virus infection and its purification to apparent homogeneity. The purified protein has an intrinsic nucleoside triphosphatase activity which is independent of, and not stimulated by, any common nucleic acid cofactors. All eight common ribo- and deoxyribonucleoside triphosphates are hydrolyzed to the diphosphate form in the presence of a divalent cation. Implications for the role of D5 in viral DNA replication are addressed.

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References

Morgan J, Cohen L, Roberts B . Identification of the DNA sequences encoding the large subunit of the mRNA-capping enzyme of vaccinia virus. J Virol. 1984; 52(1):206-14. PMC: 254507. DOI: 10.1128/JVI.52.1.206-214.1984. View

Mathews C . Enzyme organization in DNA precursor biosynthesis. Prog Nucleic Acid Res Mol Biol. 1993; 44:167-203. DOI: 10.1016/s0079-6603(08)60220-2. View

Niles E, Condit R, Caro P, Davidson K, Matusick L, Seto J . Nucleotide sequence and genetic map of the 16-kb vaccinia virus HindIII D fragment. Virology. 1986; 153(1):96-112. DOI: 10.1016/0042-6822(86)90011-5. View

Roseman N, Hruby D . Nucleotide sequence and transcript organization of a region of the vaccinia virus genome which encodes a constitutively expressed gene required for DNA replication. J Virol. 1987; 61(5):1398-406. PMC: 254115. DOI: 10.1128/JVI.61.5.1398-1406.1987. View

Evans E, Traktman P . Molecular genetic analysis of a vaccinia virus gene with an essential role in DNA replication. J Virol. 1987; 61(10):3152-62. PMC: 255892. DOI: 10.1128/JVI.61.10.3152-3162.1987. View

Broyles S, Yuen L, Shuman S, Moss B . Purification of a factor required for transcription of vaccinia virus early genes. J Biol Chem. 1988; 263(22):10754-60. View

Zhu L, Weller S . UL5, a protein required for HSV DNA synthesis: genetic analysis, overexpression in Escherichia coli, and generation of polyclonal antibodies. Virology. 1988; 166(2):366-78. DOI: 10.1016/0042-6822(88)90507-7. View

Traktman P, Kelvin M, Pacheco S . Molecular genetic analysis of vaccinia virus DNA polymerase mutants. J Virol. 1989; 63(2):841-6. PMC: 247758. DOI: 10.1128/JVI.63.2.841-846.1989. View

Elroy-Stein O, Fuerst T, Moss B . Cap-independent translation of mRNA conferred by encephalomyocarditis virus 5' sequence improves the performance of the vaccinia virus/bacteriophage T7 hybrid expression system. Proc Natl Acad Sci U S A. 1989; 86(16):6126-30. PMC: 297788. DOI: 10.1073/pnas.86.16.6126. View

10.

Gorbalenya A, Koonin E . Viral proteins containing the purine NTP-binding sequence pattern. Nucleic Acids Res. 1989; 17(21):8413-40. PMC: 335016. DOI: 10.1093/nar/17.21.8413. View

11.

Kerr S, Smith G . Vaccinia virus encodes a polypeptide with DNA ligase activity. Nucleic Acids Res. 1989; 17(22):9039-50. PMC: 335112. DOI: 10.1093/nar/17.22.9039. View

12.

Rempel R, Anderson M, Evans E, Traktman P . Temperature-sensitive vaccinia virus mutants identify a gene with an essential role in viral replication. J Virol. 1990; 64(2):574-83. PMC: 249146. DOI: 10.1128/JVI.64.2.574-583.1990. View

13.

NEUHARD J, RANDERATH E, Randerath K . Ion-exchange thin-layer chromatography. 13. Resolution of complex nucleoside triphosphate mixtures. Anal Biochem. 1965; 13(2):211-22. DOI: 10.1016/0003-2697(65)90191-0. View

14.

Siegel L, MONTY K . Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. Biochim Biophys Acta. 1966; 112(2):346-62. DOI: 10.1016/0926-6585(66)90333-5. View

15.

Paolette E, Moss B . Two nucleid acid-dependent nucleoside triphosphate phosphohydrolases from vaccinia virus. Purification and characterization. J Biol Chem. 1974; 249(10):3273-80. View

16.

Paoletti E, Moss B . Two nucleic acid-dependent nucleoside triphosphate phosphohydrolases from vaccinia virus. Nucleotide substrate and polynucleotide cofactor specificities. J Biol Chem. 1974; 249(10):3281-6. View

17.

Pelham H, Jackson R . An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976; 67(1):247-56. DOI: 10.1111/j.1432-1033.1976.tb10656.x. View

18.

Towbin H, Staehelin T, Gordon J . Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979; 76(9):4350-4. PMC: 411572. DOI: 10.1073/pnas.76.9.4350. View

19.

McFadden G, Dales S . Biogenesis of poxviruses: preliminary characterization of conditional lethal mutants of vaccinia virus defective in DNA synthesis. Virology. 1980; 103(1):68-79. DOI: 10.1016/0042-6822(80)90126-9. View

20.

Arai K, Kornberg A . Mechanism of dnaB protein action. II. ATP hydrolysis by dnaB protein dependent on single- or double-stranded DNA. J Biol Chem. 1981; 256(10):5253-9. View