Epstein-Barr virus (EBV) is a human tumor virus that is etiologically associated with nasopharyngeal and gastric cancer, Burkitt's, Hodgkin's and immunoblastic lymphoma and other human cancers. In all EBVassociated tumors and in normal B cells, the virus remains in a latent state of limited gene expression. The virus must be reactivated into its lytic cycle in order to spread between cells and among individuals. The EBV/Rta protein encoded in the viral BRLF1 gene plays an obligatory role in mediating the switch between latency and lytic cycle gene expression, acting both as a transcription factor and a replication protein. Rta is thus is a key factor in pathogenesis and oncogenesis. Our global objectives are to understand the control of BRLF1 expression, the mechanism of action of Rta and the ways Rta might manipulate expression and localization of cellular proteins. The proposed experiments are derived from many recent publications and extensive preliminary data that investigated control of expression and regulation of activity of EBV/Rta and its homologue KSHV/ORF50. We will address unsolved mysteries: Which cellular proteins regulate expression of Rta? Why do some cells respond to lytic inducing stimuli by expression of Rta while other cells are refractory? How does Rta function both as a transcription factor and replication protein? How does Rta alter the function and localization of key cellular proteins? Accordingly, our specific aims are 1) To determine the capacity and importance of cellular immediate-early proteins, such as early response and orphan nuclear receptor proteins, to activate Rta expression;2) To understand the function of Rta in EBV DNA replication, particularly the role of phosphorylation and regulation of DNA binding;and 3) To delineate the function of Rta in expression, activation and re-Iocalization of the DNA damage response cohesin, SMC1, STAT3 and PolyA binding protein. The proposed studies, utilizing tools of cell biology, biochemistry, molecular genetics, and immunology address both basic questions of control of eukaryotic gene expression that are relevant to cancer and specific unresolved issues about molecular pathogenesis of an important human cancer virus for which there is yet no specific prevention or treatment.

Public Health Relevance

Basic studies of the molecular biology of the Rta protein of Epstein-Barr virus will ultimately lead to enhanced understanding of pathogenesis and improved diagnosis, treatment and prevention of EBV-associated cancers that are of world-wide importance.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA016038-40
Application #
8476991
Study Section
Special Emphasis Panel (ZCA1-GRB-S)
Project Start
Project End
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
40
Fiscal Year
2013
Total Cost
$349,372
Indirect Cost
$135,910
Name
Yale University
Department
Type
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Zhang, Pengwei; Monteiro da Silva, Gabriel; Deatherage, Catherine et al. (2018) Cell-Penetrating Peptide Mediates Intracellular Membrane Passage of Human Papillomavirus L2 Protein to Trigger Retrograde Trafficking. Cell 174:1465-1476.e13
Inoue, Takamasa; Zhang, Pengwei; Zhang, Wei et al. (2018) ?-Secretase promotes membrane insertion of the human papillomavirus L2 capsid protein during virus infection. J Cell Biol 217:3545-3559
Vallery, Tenaya K; Withers, Johanna B; Andoh, Joana A et al. (2018) Kaposi's Sarcoma-Associated Herpesvirus mRNA Accumulation in Nuclear Foci Is Influenced by Viral DNA Replication and Viral Noncoding Polyadenylated Nuclear RNA. J Virol 92:
Hayes, Karen E; Barr, Jamie A; Xie, Mingyi et al. (2018) Immunoprecipitation of Tri-methylated Capped RNA. Bio Protoc 8:
Park, Richard; Miller, George (2018) Epstein-Barr Virus-Induced Nodules on Viral Replication Compartments Contain RNA Processing Proteins and a Viral Long Noncoding RNA. J Virol 92:
Lipovsky, Alex; Erden, Asu; Kanaya, Eriko et al. (2017) The cellular endosomal protein stannin inhibits intracellular trafficking of human papillomavirus during virus entry. J Gen Virol 98:2821-2836
Singh, Gatikrushna; Fritz, Sarah M; Ranji, Arnaz et al. (2017) Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution. J Vis Exp :
Martinez, Ivan; Hayes, Karen E; Barr, Jamie A et al. (2017) An Exportin-1-dependent microRNA biogenesis pathway during human cell quiescence. Proc Natl Acad Sci U S A 114:E4961-E4970
Pawlica, Paulina; Moss, Walter N; Steitz, Joan A (2016) Host miRNA degradation by Herpesvirus saimiri small nuclear RNA requires an unstructured interacting region. RNA 22:1181-9
Gorres, Kelly L; Daigle, Derek; Mohanram, Sudharshan et al. (2016) Valpromide Inhibits Lytic Cycle Reactivation of Epstein-Barr Virus. MBio 7:e00113

Showing the most recent 10 out of 340 publications