Herpesvirus latency depends on viral replication. Progeny viruses are needed to pass on new viral infections, and for Kaposi sarcoma-associated herpesvirus (KSHV), a certain level of replication appears to be required to maintain the latent state. Herpes simplex virus (HSV-1) in neuronal ganglia can cycle between latent and replicative states. Epstein Barr virus (EBV) infection results in mononucleosis, which provides a reservoir of latently infected cells in which the EBV genome resides as a latent episome. EBV is associated with several human cancers including nasopharyngeal carcinomas and Burkitts lymphoma. KSHV infection leads to Kaposi's sarcoma, primary effusion lymphoma, and Multicentric Castleman's disease. Our working hypothesis is that to understand the establishment and maintenance of herpesvirus latency we must understand the role of viral replication and recombination factors as well as the proteins which directly control the latent origins. Central to these pathways are the molecular complexes formed between the viral DNAs and proteins. Our efforts are focused on elucidating molecular mechanisms and structures related to viral DNA-protein interactions. This can provide insights that drive new thinking and uncover important mechanistic details.
In Aim I we will examine the structure of filaments generated by the multifunctional DNA binding proteins of HSV-1, EBV, and KSHV. Our working hypothesis is that these filaments provide structural scaffolds upon which replication and recombination occur in the infected cell nucleus and this will be tested using a combination of biochemical, genetic and and electron microscopic (EM) approaches.
In Aim II, the structure of the 2-part recombinases of the herpesviruses will be examined using biochemical and electron microscopic methods. Their action on the KSHV terminal repeats will be examined to determine if this may lead to circularization and latency.
In Aim III the binding of KSHV LANA origin binding protein together with p53 and other host proteins will be examined using EM to learn how these proteins remodel the architecture of the latent origin. These studies are highly interactive with the KSHV work being done together with the Dittmer and Damania laboratories.

Public Health Relevance

These studies will generate unique the structural and biochemical information needed to provide the basis for a new understanding of the molecular basis for latency and lytic replication in the herpesviruses at the level of DNA-protein complexes. These results will, in turn, provide new inroads and thinking that are needed to generate anti-viral approaches.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
2P01CA019014-32A1
Application #
8166052
Study Section
Special Emphasis Panel (ZCA1-RPRB-0 (M1))
Project Start
1997-05-01
Project End
2016-06-30
Budget Start
2011-07-27
Budget End
2012-06-30
Support Year
32
Fiscal Year
2011
Total Cost
$270,533
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Bigi, Rachele; Landis, Justin T; An, Hyowon et al. (2018) Epstein-Barr virus enhances genome maintenance of Kaposi sarcoma-associated herpesvirus. Proc Natl Acad Sci U S A 115:E11379-E11387
El-Mallawany, Nader Kim; Villiera, Jimmy; Kamiyango, William et al. (2018) Endemic Kaposi sarcoma in HIV-negative children and adolescents: an evaluation of overlapping and distinct clinical features in comparison with HIV-related disease. Infect Agent Cancer 13:33
Kobayashi, E; Aga, M; Kondo, S et al. (2018) C-Terminal Farnesylation of UCH-L1 Plays a Role in Transport of Epstein-Barr Virus Primary Oncoprotein LMP1 to Exosomes. mSphere 3:
Hopcraft, Sharon E; Pattenden, Samantha G; James, Lindsey I et al. (2018) Chromatin remodeling controls Kaposi's sarcoma-associated herpesvirus reactivation from latency. PLoS Pathog 14:e1007267
Sin, Sang-Hoon; Eason, Anthony B; Bigi, Rachele et al. (2018) Kaposi's Sarcoma-Associated Herpesvirus Latency Locus Renders B Cells Hyperresponsive to Secondary Infections. J Virol 92:
Zhang, Yugen; Dittmer, Dirk P; Mieczkowski, Piotr A et al. (2018) RIG-I Detects Kaposi's Sarcoma-Associated Herpesvirus Transcripts in a RNA Polymerase III-Independent Manner. MBio 9:
Anders, Penny M; Montgomery, Nathan D; Montgomery, Stephanie A et al. (2018) Human herpesvirus-encoded kinase induces B cell lymphomas in vivo. J Clin Invest 128:2519-2534
Ciesielski, Grzegorz L; Nadalutti, Cristina A; Oliveira, Marcos T et al. (2018) Structural rearrangements in the mitochondrial genome of Drosophila melanogaster induced by elevated levels of the replicative DNA helicase. Nucleic Acids Res 46:3034-3046
Gurung, Sunam; Preno, Alisha N; Dubaut, Jamie P et al. (2018) Translational Model of Zika Virus Disease in Baboons. J Virol :
McNamara, Ryan P; Costantini, Lindsey M; Myers, T Alix et al. (2018) Nef Secretion into Extracellular Vesicles or Exosomes Is Conserved across Human and Simian Immunodeficiency Viruses. MBio 9:

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