The long-term goals of this research are to define the mechanisms of action of the herpes simplex virus ICP8 ssDNA binding protein in viral DNA replication and late gene transcription and the mechanisms of assembly and maturation of viral replication compartments in the infected cell nucleus. In this application our specific aims are to: 1. Define the mechanisms of stimulation of late gene expression by ICP8 by: determining if ICP8 and ICP27 interact directly by studying in vitro expressed proteins, baculovirus expressed proteins, and fluorescence resonance energy transfer analysis, by mapping the portions of ICP8 that are needed for interaction with ICP27 and pol II holoenzyme, by determining if viral DNA synthesis and ICP8 bound to progeny viral DNA are needed continuously for late transcription, by performing chromatin immunoprecipitation studies to determine if ICP8 is needed to recruit Pol II to progeny viral DNA for late transcription. 2. Define the mechanisms of intranuclear movement of ICP8 and the mechanisms of formation of replication compartments by mapping the ICP8 sequences needed for movement to prereplicative sites and replication compartments in the infected cell nucleus, by defining the domains of ICP0 needed for promotion of replication compartment formation, by testing the need for metabolic energy and actin-based myosin activity for movement of ICP8 within the nucleus. 3. Determine the mechanisms by which the HSV UL31 and UL34 gene products promote late replication compartment maturation through the host chromatin and nuclear lamina by testing the hypothesis that UL31 and UL34 cause the phosphorylation of the lamin proteins, by testing the hypothesis that UL31 binds to lamins and ? causes their disaggregation, and by determining other proteins associated with UL31 and UL34 as candidates for effectors in nuclear structural changes. These studies are likely to provide insight into the mechanisms by which HSV assembles a new apparatus for viral DNA synthesis, late gene transcription, capsid assembly, and primary envelopment in the infected cell nucleus as well as potentially providing new targets for prevention and treatment of herpetic infections. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI063106-23
Application #
6921901
Study Section
Virology Study Section (VR)
Program Officer
Beisel, Christopher E
Project Start
1979-08-01
Project End
2009-01-31
Budget Start
2005-02-01
Budget End
2006-01-31
Support Year
23
Fiscal Year
2005
Total Cost
$381,375
Indirect Cost
Name
Harvard University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
047006379
City
Boston
State
MA
Country
United States
Zip Code
02115
Peng, Tao; Chanthaphavong, R Savanh; Sun, Sijie et al. (2017) Keratinocytes produce IL-17c to protect peripheral nervous systems during human HSV-2 reactivation. J Exp Med 214:2315-2329
Hendricks, Gabriel L; Velazquez, Lourdes; Pham, Serena et al. (2015) Heparin octasaccharide decoy liposomes inhibit replication of multiple viruses. Antiviral Res 116:34-44
Suk, Hyung; Knipe, David M (2015) Proteomic analysis of the herpes simplex virus 1 virion protein 16 transactivator protein in infected cells. Proteomics 15:1957-67
Knipe, David M (2015) Nuclear sensing of viral DNA, epigenetic regulation of herpes simplex virus infection, and innate immunity. Virology 479-480:153-9
Oh, Hyung Suk; Traktman, Paula; Knipe, David M (2015) Barrier-to-Autointegration Factor 1 (BAF/BANF1) Promotes Association of the SETD1A Histone Methyltransferase with Herpes Simplex Virus Immediate-Early Gene Promoters. MBio 6:e00345-15
Deng, Zhong; Kim, Eui Tae; Vladimirova, Olga et al. (2014) HSV-1 remodels host telomeres to facilitate viral replication. Cell Rep 9:2263-78
Bonham, Kevin S; Orzalli, Megan H; Hayashi, Kachiko et al. (2014) A promiscuous lipid-binding protein diversifies the subcellular sites of toll-like receptor signal transduction. Cell 156:705-16
Oh, Hyung Suk; Bryant, Kevin F; Nieland, Thomas J F et al. (2014) A targeted RNA interference screen reveals novel epigenetic factors that regulate herpesviral gene expression. MBio 5:e01086-13
Yan, Zhipeng; Bryant, Kevin F; Gregory, Sean M et al. (2014) HIV integrase inhibitors block replication of alpha-, beta-, and gammaherpesviruses. MBio 5:e01318-14
Larson, Alyssa M; Oh, Hyung Suk; Knipe, David M et al. (2013) Decreasing herpes simplex viral infectivity in solution by surface-immobilized and suspended N,N-dodecyl,methyl-polyethylenimine. Pharm Res 30:25-31

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