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Animal: DNA Viruses |
MRC Virology Unit, Institute of Virology, Church Street, Glasgow G11 5JR, UK1
Author for correspondence: Nigel Stow. Fax +44 141 337 2236. e-mail n.stow{at}vir.gla.ac.uk
The UL15 and UL28 proteins of herpes simplex virus type 1 are both required for the packaging of replicated viral DNA into the viral capsid. We have expressed UL28 and a functional epitope-tagged form of UL15 in mammalian and insect cells. Immunoprecipitation experiments confirmed that the two proteins can interact. In agreement with previous results, UL15, when expressed alone, entered the nucleus but UL28 remained cytoplasmic. When co-expressed the two proteins co-localized in the nucleus. Six UL28 deletion mutants were constructed and similarly analysed. The results obtained by immunoprecipitation and immunofluorescence were consistent and demonstrate that at least two separate regions of the UL28 polypeptide chain have the ability to interact with UL15. Surprisingly, three of the mutants prevented the UL15 protein from localizing to the cell nucleus, and these were not functional in a transient DNA packaging assay. Of the three UL28 mutant proteins that entered the nucleus with UL15, one containing an internal deletion of 13 amino acids was able to complement a UL28 null mutant in both DNA packaging and virus yield assays, demonstrating that this region of the protein is not essential for function. In addition to interacting with the UL28 protein we also demonstrated that UL15 molecules can interact with each other, and that sequences within the second exon contribute to this interaction.
This article has been cited by other articles:
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E. Wills, L. Scholtes, and J. D. Baines Herpes Simplex Virus 1 DNA Packaging Proteins Encoded by UL6, UL15, UL17, UL28, and UL33 Are Located on the External Surface of the Viral Capsid J. Virol., November 1, 2006; 80(21): 10894 - 10899. [Abstract] [Full Text] [PDF]
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K. Yang and J. D. Baines The Putative Terminase Subunit of Herpes Simplex Virus 1 Encoded by UL28 Is Necessary and Sufficient To Mediate Interaction between pUL15 and pUL33. J. Virol., June 1, 2006; 80(12): 5733 - 5739. [Abstract] [Full Text] [PDF]
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M. A. McVoy and D. E. Nixon Impact of 2-Bromo-5,6-Dichloro-1-{beta}-D-Ribofuranosyl Benzimidazole Riboside and Inhibitors of DNA, RNA, and Protein Synthesis on Human Cytomegalovirus Genome Maturation J. Virol., September 1, 2005; 79(17): 11115 - 11127. [Abstract] [Full Text] [PDF]
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I. M. Porter and N. D. Stow Replication, recombination and packaging of amplicon DNA in cells infected with the herpes simplex virus type 1 alkaline nuclease null mutant ambUL12 J. Gen. Virol., December 1, 2004; 85(12): 3501 - 3510. [Abstract] [Full Text] [PDF]
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D. E. Nixon and M. A. McVoy Dramatic Effects of 2-Bromo-5,6-Dichloro-1-{beta}-D-Ribofuranosyl Benzimidazole Riboside on the Genome Structure, Packaging, and Egress of Guinea Pig Cytomegalovirus J. Virol., February 15, 2004; 78(4): 1623 - 1635. [Abstract] [Full Text] [PDF]
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P. M. Beard, C. Duffy, and J. D. Baines Quantification of the DNA Cleavage and Packaging Proteins UL15 and UL28 in A and B Capsids of Herpes Simplex Virus Type 1 J. Virol., February 1, 2004; 78(3): 1367 - 1374. [Abstract] [Full Text] [PDF]
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A. J. Przech, D. Yu, and S. K. Weller Point Mutations in Exon I of the Herpes Simplex Virus Putative Terminase Subunit, UL15, Indicate that the Most Conserved Residues Are Essential for Cleavage and Packaging J. Virol., September 1, 2003; 77(17): 9613 - 9621. [Abstract] [Full Text] [PDF]
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C. A. White, N. D. Stow, A. H. Patel, M. Hughes, and V. G. Preston Herpes Simplex Virus Type 1 Portal Protein UL6 Interacts with the Putative Terminase Subunits UL15 and UL28 J. Virol., June 1, 2003; 77(11): 6351 - 6358. [Abstract] [Full Text] [PDF]
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V. Ellsmore, G. G. Reid, and N. D. Stow Detection of human cytomegalovirus DNA replication in non-permissive Vero and 293 cells J. Gen. Virol., March 1, 2003; 84(3): 639 - 645. [Abstract] [Full Text] [PDF]
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P. M. Beard, N. S. Taus, and J. D. Baines DNA Cleavage and Packaging Proteins Encoded by Genes UL28, UL15, and UL33 of Herpes Simplex Virus Type 1 Form a Complex in Infected Cells J. Virol., April 16, 2002; 76(10): 4785 - 4791. [Abstract] [Full Text] [PDF]
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 H. Scheffczik, C. G. W. Savva, A. Holzenburg, L. Kolesnikova, and E. Bogner The terminase subunits pUL56 and pUL89 of human cytomegalovirus are DNA-metabolizing proteins with toroidal structure Nucleic Acids Res., April 1, 2002; 30(7): 1695 - 1703. [Abstract] [Full Text] [PDF]
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N. D. Stow Packaging of Genomic and Amplicon DNA by the Herpes Simplex Virus Type 1 UL25-Null Mutant KUL25NS J. Virol., November 15, 2001; 75(22): 10755 - 10765. [Abstract] [Full Text] [PDF]
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P. D. Hodge and N. D. Stow Effects of Mutations within the Herpes Simplex Virus Type 1 DNA Encapsidation Signal on Packaging Efficiency J. Virol., October 1, 2001; 75(19): 8977 - 8986. [Abstract] [Full Text] [PDF]
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