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Journal of Virology, August 2007, p. 8742-8751, Vol. 81, No. 16
0022-538X/07/$08.00+0     doi:10.1128/JVI.00174-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

A Mutation in the Human Herpes Simplex Virus Type 1 UL52 Zinc Finger Motif Results in Defective Primase Activity but Can Recruit Viral Polymerase and Support Viral Replication Efficiently{triangledown}

Yan Chen, Christine M. Livingston, Stacy D. Carrington-Lawrence,{dagger} Ping Bai, and Sandra K. Weller*

Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, Connecticut 06030

Received 25 January 2007/ Accepted 21 May 2007

Herpes simplex virus type 1 (HSV-1) encodes a heterotrimeric helicase/primase complex consisting of UL5, UL8, and UL52. UL5 contains conserved helicase motifs, while UL52 contains conserved primase motifs, including a zinc finger motif. Although HSV-1 and HSV-2 UL52s contain a leucine residue at position 986, most other herpesvirus primase homologues contain a phenylalanine at this position. We constructed an HSV-1 UL52 L986F mutation and found that it can complement a UL52 null virus more efficiently than the wild type (WT). We thus predicted that the UL5/8/52 complex containing the L986F mutation might posses increased primase activity; however, it exhibited only 25% of the WT level of primase activity. Interestingly, the mutant complex displayed elevated levels of DNA binding and single-stranded DNA-dependent ATPase and helicase activities. This result confirms a complex interdependence between the helicase and primase subunits. We previously showed that primase-defective mutants failed to recruit the polymerase catalytic subunit UL30 to prereplicative sites, suggesting that an active primase, or primer synthesis, is required for polymerase recruitment. Although L986F exhibits decreased primase activity, it can support efficient replication and recruit UL30 efficiently to replication compartments, indicating that a partially active primase is capable of recruiting polymerase. Extraction with detergents prior to fixation can extract nucleosolic proteins but not proteins bound to chromatin or the nuclear matrix. We showed that UL30 was extracted from replication compartments while UL42 remained bound, suggesting that UL30 may be tethered to the replication fork by protein-protein interactions.

* Corresponding author. Mailing address: Department of Molecular, Microbial and Structural Biology, MC3205, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030. Phone: (860) 679-2310. Fax: (860) 679-1239. E-mail: weller{at}nso2.uchc.edu

{triangledown} Published ahead of print on 6 June 2007.

{dagger} Current address: National Institutes of Health, Office of the Director, 1 Center Drive, Bethesda, MD 20892.

Journal of Virology, August 2007, p. 8742-8751, Vol. 81, No. 16
0022-538X/07/$08.00+0     doi:10.1128/JVI.00174-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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  • Biswas, S., Kleymann, G., Swift, M., Tiley, L. S., Lyall, J., Aguirre-Hernandez, J., Field, H. J. (2008). A single drug-resistance mutation in HSV-1 UL52 primase points to a difference between two helicase-primase inhibitors in their mode of interaction with the antiviral target. J Antimicrob Chemother 61: 1044-1047 [Abstract] [Full Text]  


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