This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Uprichard, S. L. Articles by Knipe, D. M. Search for Related Content PubMed PubMed Citation Articles by Uprichard, S. L. Articles by Knipe, D. M.

 Previous Article  |  Next Article 

Journal of Virology, July 2003, p. 7467-7476, Vol. 77, No. 13
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.13.7467-7476.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Conformational Changes in the Herpes Simplex Virus ICP8 DNA-Binding Protein Coincident with Assembly in Viral Replication Structures

Susan L. Uprichard and David M. Knipe^*

Committee on Virology and Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

Received 24 January 2003/ Accepted 10 April 2003

The herpes simplex virus (HSV) single-stranded DNA-binding protein, ICP8, is required for viral DNA synthesis. Before viral DNA replication, ICP8 colocalizes with other replication proteins at small punctate foci called prereplicative sites. With the onset of viral genome amplification, these proteins become redistributed into large globular replication compartments. Here we present the results of immunocytochemical and biochemical analysis of ICP8 showing that various antibodies recognize distinct forms of ICP8. Using these ICP8-specific antibodies as probes for ICP8 structure, we detected a time-dependent appearance and disappearance of ICP8 epitopes in immunoprecipitation assays. Immunofluorescence staining of ICP8 in cells infected with different HSV mutant viruses as well as cells transfected with a limited number of viral genes demonstrated that these and other antigenic changes occur coincident with ICP8 assembly at intranuclear replication structures. Genetic analysis has revealed a correlation between the ability of various ICP8 mutant proteins to form the 39S epitope and their ability to bind to DNA. These results support the hypothesis that ICP8 undergoes a conformational change upon binding to other HSV proteins and/or to DNA coincident with assembly into viral DNA replication structures.

---

* Corresponding author. Mailing address: Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-1934. Fax: (617) 432-0223. E-mail: david_knipe{at}hms.harvard.edu.

Present address: The Scripps Research Institute, La Jolla, CA 92037.

---

Journal of Virology, July 2003, p. 7467-7476, Vol. 77, No. 13
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.13.7467-7476.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Su, Y.-H., Zhang, X., Wang, X., Fraser, N. W., Block, T. M. (2006). Evidence that the Immediate-Early Gene Product ICP4 Is Necessary for the Genome of the Herpes Simplex Virus Type 1 ICP4 Deletion Mutant Strain d120 To Circularize in Infected Cells. J. Virol. 80: 11589-11597 [Abstract] [Full Text]  
• Mikhailov, V. S., Okano, K., Rohrmann, G. F. (2005). The Redox State of the Baculovirus Single-stranded DNA-binding Protein LEF-3 Regulates Its DNA Binding, Unwinding, and Annealing Activities. J. Biol. Chem. 280: 29444-29453 [Abstract] [Full Text]  
• Taylor, T. J., Knipe, D. M. (2004). Proteomics of Herpes Simplex Virus Replication Compartments: Association of Cellular DNA Replication, Repair, Recombination, and Chromatin Remodeling Proteins with ICP8. J. Virol. 78: 5856-5866 [Abstract] [Full Text]