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

 Previous Article  |  Next Article 

Journal of Virology, November 1998, p. 8772-8781, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

In Vitro Processing of Herpes Simplex Virus Type 1 DNA Replication Intermediates by the Viral Alkaline Nuclease, UL12

Joshua N. Goldstein and Sandra K. Weller*

Department of Microbiology, University of Connecticut Health Center, Farmington, Connecticut 06030

Received 26 May 1998/Accepted 11 August 1998

Herpes simplex virus type 1 (HSV-1) DNA replication intermediates exist in a complex nonlinear structure that does not migrate into a pulsed-field gel. Genetic evidence suggests that the product of the UL12 gene, termed alkaline nuclease, plays a role in processing replication intermediates (R. Martinez, R. T. Sarisky, P. C. Weber, and S. K. Weller, J. Virol. 70:2075-2085, 1996). In this study we have tested the hypothesis that alkaline nuclease acts as a structure-specific resolvase. Cruciform structures generated with oligonucleotides were treated with purified alkaline nuclease; however, instead of being resolved into linear duplexes as would be expected of a resolvase activity, the artificial cruciforms were degraded. DNA replication intermediates were isolated from the well of a pulsed-field gel ("well DNA") and treated with purified HSV-1 alkaline nuclease. Although alkaline nuclease can degrade virion DNA to completion, digestion of well DNA results in a smaller-than-unit-length product that migrates as a heterogeneous smear; this product is resistant to further digestion by alkaline nuclease. The smaller-than-unit-length products are representative of the entire HSV genome, indicating that alkaline nuclease is not inhibited at specific sequences. To further probe the structure of replicating DNA, well DNA was treated with various known nucleases; our results indicate that replicating DNA apparently contains no accessible double-stranded ends but does contain nicks and gaps. Our data suggest that UL12 functions at nicks and gaps in replicating DNA to correctly repair or process the replicating genome into a form suitable for encapsidation.

* Corresponding author. Mailing address: MC-3205, Department of Microbiology, University of Connecticut Health Center, Farmington, CT 06030-3205. Phone: (860) 679-2310. Fax: (860) 679-1239. E-mail: weller{at}nso2.uchc.edu.

Journal of Virology, November 1998, p. 8772-8781, Vol. 72, No. 11
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Covarrubias, S., Richner, J. M., Clyde, K., Lee, Y. J., Glaunsinger, B. A. (2009). Host Shutoff Is a Conserved Phenotype of Gammaherpesvirus Infection and Is Orchestrated Exclusively from the Cytoplasm. J. Virol. 83: 9554-9566 [Abstract] [Full Text]  
  • Feederle, R., Bannert, H., Lips, H., Muller-Lantzsch, N., Delecluse, H.-J. (2009). The Epstein-Barr Virus Alkaline Exonuclease BGLF5 Serves Pleiotropic Functions in Virus Replication. J. Virol. 83: 4952-4962 [Abstract] [Full Text]  
  • Corcoran, J. A., Saffran, H. A., Duguay, B. A., Smiley, J. R. (2009). Herpes Simplex Virus UL12.5 Targets Mitochondria through a Mitochondrial Localization Sequence Proximal to the N Terminus. J. Virol. 83: 2601-2610 [Abstract] [Full Text]  
  • Burch, A. D., Weller, S. K. (2005). Herpes Simplex Virus Type 1 DNA Polymerase Requires the Mammalian Chaperone Hsp90 for Proper Localization to the Nucleus. J. Virol. 79: 10740-10749 [Abstract] [Full Text]  
  • Chen, Y., Carrington-Lawrence, S. D., Bai, P., Weller, S. K. (2005). Mutations in the Putative Zinc-Binding Motif of UL52 Demonstrate a Complex Interdependence between the UL5 and UL52 Subunits of the Human Herpes Simplex Virus Type 1 Helicase/Primase Complex. J. Virol. 79: 9088-9096 [Abstract] [Full Text]  
  • Glaunsinger, B., Chavez, L., Ganem, D. (2005). The Exonuclease and Host Shutoff Functions of the SOX Protein of Kaposi's Sarcoma-Associated Herpesvirus Are Genetically Separable. J. Virol. 79: 7396-7401 [Abstract] [Full Text]  
  • Porter, I. M., Stow, N. D. (2004). 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. 85: 3501-3510 [Abstract] [Full Text]  
  • Okano, K., Vanarsdall, A. L., Rohrmann, G. F. (2004). Characterization of a Baculovirus Lacking the Alkaline Nuclease Gene. J. Virol. 78: 10650-10656 [Abstract] [Full Text]  
  • Mikhailov, V. S., Okano, K., Rohrmann, G. F. (2004). Specificity of the Endonuclease Activity of the Baculovirus Alkaline Nuclease for Single-stranded DNA. J. Biol. Chem. 279: 14734-14745 [Abstract] [Full Text]  
  • Mikhailov, V. S., Okano, K., Rohrmann, G. F. (2003). Baculovirus Alkaline Nuclease Possesses a 5'->3' Exonuclease Activity and Associates with the DNA-Binding Protein LEF-3. J. Virol. 77: 2436-2444 [Abstract] [Full Text]  
  • Umene, K. (2001). Cleavage in and around the DR1 Element of the a Sequence of Herpes Simplex Virus Type 1 Relevant to the Excision of DNA Fragments with Length Corresponding to One and Two Units of the a Sequence. J. Virol. 75: 5870-5878 [Abstract] [Full Text]  
  • Aravind, L., Makarova, K. S., Koonin, E. V. (2000). SURVEY AND SUMMARY: Holliday junction resolvases and related nucleases: identification of new families, phyletic distribution and evolutionary trajectories. Nucleic Acids Res 28: 3417-3432 [Abstract] [Full Text]  
  • Li, L., Rohrmann, G. F. (2000). Characterization of a Baculovirus Alkaline Nuclease. J. Virol. 74: 6401-6407 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»