Abstract
We used indirect immunofluorescence to examine the factors determining the intranuclear location of herpes simplex virus (HSV) DNA polymerase (Pol) in infected cells. In the absence of viral DNA replication, HSV Pol colocalized with the HSV DNA-binding protein ICP8 in nuclear framework-associated structures called prereplicative sites. In the presence of viral DNA replication, HSV Pol colocalized with ICP8 in globular intranuclear structures called replication compartments. In cells infected with mutant viruses encoding defective ICP8 molecules, Pol localized within the cell nucleus but showed a general diffuse intranuclear distribution. In uninfected cells transfected with a plasmid expressing Pol, Pol similarly showed a diffuse intranuclear distribution. Therefore, Pol can localize to the cell nucleus without other viral proteins, but functional ICP8 is required for Pol to localize to prereplicative sites. In cells infected with mutant viruses encoding defective Pol molecules, ICP8 localized to prereplicative sites. Thus, Pol or the portions of Pol not expressed by the mutant viruses are not essential for the formation of prereplicative sites or the localization of ICP8 to these structures. These results demonstrate that a specific nuclear protein can influence the intranuclear location of another nuclear protein.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Challberg M. D., Kelly T. J. Animal virus DNA replication. Annu Rev Biochem. 1989;58:671–717. doi: 10.1146/annurev.bi.58.070189.003323. [DOI] [PubMed] [Google Scholar]
- Chiou H. C., Weller S. K., Coen D. M. Mutations in the herpes simplex virus major DNA-binding protein gene leading to altered sensitivity to DNA polymerase inhibitors. Virology. 1985 Sep;145(2):213–226. doi: 10.1016/0042-6822(85)90155-2. [DOI] [PubMed] [Google Scholar]
- Gao M., Knipe D. M. Genetic evidence for multiple nuclear functions of the herpes simplex virus ICP8 DNA-binding protein. J Virol. 1989 Dec;63(12):5258–5267. doi: 10.1128/jvi.63.12.5258-5267.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hernandez T. R., Lehman I. R. Functional interaction between the herpes simplex-1 DNA polymerase and UL42 protein. J Biol Chem. 1990 Jul 5;265(19):11227–11232. [PubMed] [Google Scholar]
- Knipe D. M., Senechek D., Rice S. A., Smith J. L. Stages in the nuclear association of the herpes simplex virus transcriptional activator protein ICP4. J Virol. 1987 Feb;61(2):276–284. doi: 10.1128/jvi.61.2.276-284.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knipe D. M., Smith J. L. A mutant herpesvirus protein leads to a block in nuclear localization of other viral proteins. Mol Cell Biol. 1986 Jul;6(7):2371–2381. doi: 10.1128/mcb.6.7.2371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knipe D. M. The role of viral and cellular nuclear proteins in herpes simplex virus replication. Adv Virus Res. 1989;37:85–123. doi: 10.1016/s0065-3527(08)60833-7. [DOI] [PubMed] [Google Scholar]
- Leinbach S. S., Reno J. M., Lee L. F., Isbell A. F., Boezi J. A. Mechanism of phosphonoacetate inhibition of herpesvirus-induced DNA polymerase. Biochemistry. 1976 Jan 27;15(2):426–430. doi: 10.1021/bi00647a029. [DOI] [PubMed] [Google Scholar]
- Littler E., Purifoy D., Minson A., Powell K. L. Herpes simplex virus non-structural proteins. III. Function of the major DNA-binding protein. J Gen Virol. 1983 May;64(Pt 5):983–995. doi: 10.1099/0022-1317-64-5-983. [DOI] [PubMed] [Google Scholar]
- Mao J. C., Robishaw E. E. Mode of inhibition of herpes simplex virus DNA polymerase by phosphonoacetate. Biochemistry. 1975 Dec 16;14(25):5475–5479. doi: 10.1021/bi00696a015. [DOI] [PubMed] [Google Scholar]
- Marcy A. I., Yager D. R., Coen D. M. Isolation and characterization of herpes simplex virus mutants containing engineered mutations at the DNA polymerase locus. J Virol. 1990 May;64(5):2208–2216. doi: 10.1128/jvi.64.5.2208-2216.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- O'Donnell M. E., Elias P., Funnell B. E., Lehman I. R. Interaction between the DNA polymerase and single-stranded DNA-binding protein (infected cell protein 8) of herpes simplex virus 1. J Biol Chem. 1987 Mar 25;262(9):4260–4266. [PubMed] [Google Scholar]
- Olivo P. D., Nelson N. J., Challberg M. D. Herpes simplex virus type 1 gene products required for DNA replication: identification and overexpression. J Virol. 1989 Jan;63(1):196–204. doi: 10.1128/jvi.63.1.196-204.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Quinlan M. P., Chen L. B., Knipe D. M. The intranuclear location of a herpes simplex virus DNA-binding protein is determined by the status of viral DNA replication. Cell. 1984 Apr;36(4):857–868. doi: 10.1016/0092-8674(84)90035-7. [DOI] [PubMed] [Google Scholar]
- Randall R. E., Dinwoodie N. Intranuclear localization of herpes simplex virus immediate-early and delayed-early proteins: evidence that ICP 4 is associated with progeny virus DNA. J Gen Virol. 1986 Oct;67(Pt 10):2163–2177. doi: 10.1099/0022-1317-67-10-2163. [DOI] [PubMed] [Google Scholar]
- Rixon F. J., Atkinson M. A., Hay J. Intranuclear distribution of herpes simplex virus type 2 DNA synthesis: examination by light and electron microscopy. J Gen Virol. 1983 Sep;64(Pt 9):2087–2092. doi: 10.1099/0022-1317-64-9-2087. [DOI] [PubMed] [Google Scholar]
- Ruyechan W. T., Weir A. C. Interaction with nucleic acids and stimulation of the viral DNA polymerase by the herpes simplex virus type 1 major DNA-binding protein. J Virol. 1984 Dec;52(3):727–733. doi: 10.1128/jvi.52.3.727-733.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Showalter S. D., Zweig M., Hampar B. Monoclonal antibodies to herpes simplex virus type 1 proteins, including the immediate-early protein ICP 4. Infect Immun. 1981 Dec;34(3):684–692. doi: 10.1128/iai.34.3.684-692.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas M. S., Banks L. M., Purifoy D. J., Powell K. L. Production of antibodies of predetermined specificity against herpes simplex virus DNA polymerase and their use in characterization of the enzyme. J Virol. 1988 May;62(5):1550–1557. doi: 10.1128/jvi.62.5.1550-1557.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weisshart K., Knopf C. W. The herpes simplex virus type I DNA polymerase. Polypeptide structure and antigenic domains. Eur J Biochem. 1988 Jul 1;174(4):707–716. doi: 10.1111/j.1432-1033.1988.tb14155.x. [DOI] [PubMed] [Google Scholar]
- Yager D. R., Coen D. M. Analysis of the transcript of the herpes simplex virus DNA polymerase gene provides evidence that polymerase expression is inefficient at the level of translation. J Virol. 1988 Jun;62(6):2007–2015. doi: 10.1128/jvi.62.6.2007-2015.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yager D. R., Marcy A. I., Coen D. M. Translational regulation of herpes simplex virus DNA polymerase. J Virol. 1990 May;64(5):2217–2225. doi: 10.1128/jvi.64.5.2217-2225.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Bruyn Kops A., Knipe D. M. Formation of DNA replication structures in herpes virus-infected cells requires a viral DNA binding protein. Cell. 1988 Dec 2;55(5):857–868. doi: 10.1016/0092-8674(88)90141-9. [DOI] [PubMed] [Google Scholar]