The Herpesviridae are responsible for debilitating acute and congenital infections;furthermore, some members of this family are also associated with human cancers. Human Herpes Simplex Viruses Types 1 and 2 (HSV-1 and HSV-2) are responsible for primary and recurrent herpetic lesions of the mouth and genital tract as well as more serious and potentially life-threatening infections of the eye and central nervous system. HSV infections are of particular concern in newborns and in immunocompromised patients. The herpesviruses encode a large number of replication proteins that are excellent targets for antiviral therapy;however, the mechanisms by which they function during DNA replication are poorly understood. The experiments described in this proposal will focus on the interactions of these replication proteins with one another, with DNA at the replication fork and with cellular proteins that may play roles in this process. Events leading to the initiation of viral replication at the origins of replication, the formation of a multiprotein complex at the viral replisome, the priming of Okazaki fragments and interactions with cellular factors during HSV DNA replication will be examined using molecular genetic, biochemical, biophysical and cell biological approaches. Although these features of DNA replication are common for all life forms using DNA as their genetic material, HSV is an ideal system to examine them at the molecular level because this system is amenable to both genetic and biochemical manipulation. The molecular and biochemical characterization of HSV DNA replication is expected to facilitate our understanding of analogous processes in other viral and eukaryotic systems as well as to provide novel targets for antiviral therapy.
The specific aims of this proposal are:
Aim 1. How is DNA replication initiated in HSV-1 infected cells? Aim 2. How are viral and cellular proteins recruited into multiprotein assemblies that make up the replisome? Aim 3. Identify subdomains of HSV replication proteins for the purpose of domain mapping and eventual structural analysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021747-28
Application #
8215816
Study Section
Special Emphasis Panel (ZRG1-IDM-M (04))
Program Officer
Challberg, Mark D
Project Start
1984-12-01
Project End
2013-11-30
Budget Start
2012-03-01
Budget End
2013-11-30
Support Year
28
Fiscal Year
2012
Total Cost
$536,513
Indirect Cost
$174,004
Name
University of Connecticut
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
022254226
City
Farmington
State
CT
Country
United States
Zip Code
06030
Weller, Sandra K; Sawitzke, James A (2014) Recombination promoted by DNA viruses: phage ? to herpes simplex virus. Annu Rev Microbiol 68:237-58
Grady, Lorry M; Bai, Ping; Weller, Sandra K (2014) HSV-1 protein expression using recombinant baculoviruses. Methods Mol Biol 1144:293-304
Smith, Samantha; Reuven, Nina; Mohni, Kareem N et al. (2014) Structure of the herpes simplex virus 1 genome: manipulation of nicks and gaps can abrogate infectivity and alter the cellular DNA damage response. J Virol 88:10146-56
Mohni, Kareem N; Dee, Alexander R; Smith, Samantha et al. (2013) Efficient herpes simplex virus 1 replication requires cellular ATR pathway proteins. J Virol 87:531-42
Mohni, Kareem N; Smith, Samantha; Dee, Alexander R et al. (2013) Herpes simplex virus type 1 single strand DNA binding protein and helicase/primase complex disable cellular ATR signaling. PLoS Pathog 9:e1003652
Weller, Sandra K; Kuchta, Robert D (2013) The DNA helicase-primase complex as a target for herpes viral infection. Expert Opin Ther Targets 17:1119-32
Mohni, Kareem N; Mastrocola, Adam S; Bai, Ping et al. (2011) DNA mismatch repair proteins are required for efficient herpes simplex virus 1 replication. J Virol 85:12241-53
Chen, Yan; Bai, Ping; Mackay, Shannon et al. (2011) Herpes simplex virus type 1 helicase-primase: DNA binding and consequent protein oligomerization and primase activation. J Virol 85:968-78
Weller, Sandra K (2010) Herpes simplex virus reorganizes the cellular DNA repair and protein quality control machinery. PLoS Pathog 6:e1001105
Nicolas, Armel; Alazard-Dany, Nathalie; Biollay, Coline et al. (2010) Identification of rep-associated factors in herpes simplex virus type 1-induced adeno-associated virus type 2 replication compartments. J Virol 84:8871-87

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