The roles of noncoding (nc)RNAs in lymphoid cells harboring one of three transforming herpesviruses are being investigated. Epstein-Barr virus (EBV) infects and transforms human B cells; it is the causative agent of infectious mononucleosis and is associated with several human cancers. Herpesvirus saimiri (HVS) induces fatal lymphomas and leukemias in New World monkeys and transforms monkey T lymphocytes in culture. Kaposi's sarcoma-associated herpesvirus (KSHV) afflicts immunocompromised individuals and persists in a latent form until lytic activation. The two EBV-encoded EBERs, the newly discovered EBV stable intronic sequence (sis)RNA, seven HVS-encoded HSURs, and six HVS microRNAs are all expressed in virally transformed cells. Upon induction, KSHV produces PAN, a capped, polyadenylated nuclear RNA discovered in George Miller's laboratory. These viral ncRNAs are abundant, conserved and bind host proteins. Our functional studies have uncovered novel mechanisms of microRNA biogenesis and decay, identified the role of triple helices in RNA stabilization, and contributed important insights into viral evolution and host pathways perturbed during viral latency or lytic growth. Proposed aims will extend these advances to further elucidate molecular mechanisms. The contributions of HSURs1 and 2 and HVS microRNAs to T-cell activation, proliferation and immune surveillance are being validated. HVS microRNA processing will be scrutinized for the novel use of the host Integrator complex. Since PAN RNA is essential for KSHV late gene expression, we will ask whether its interaction with host poly(A) binding protein C1 (PABPC1) relocalized to the nucleus during lytic infection is essential and whether late viral mRNAs escape host shutoff by exiting the nucleus via a novel pathway involving nuclear envelope budding. Global approaches to EBER function have implicated them in post-transcriptional events in gene expression. EBER2 RNPs have been localized, along with the important B-cell transcription factor Pax5, at the terminal repeats of EBV episomes, suggesting a role in the maintenance of latency. The EBV sisRNA-1 is predicted to bind host microRNAs, leading to their sequestration or degradation. Structural studies of the KSHV PAN RNA stability element complexed with PABPC1 protein will provide molecular insights.

Public Health Relevance

Project 3, Narrative We are studying how herpesviruses that cause lymphomas or sarcomas in humans or monkeys use RNA molecules to manipulate their host cells to the advantage of the virus. These RNAs are not messages for proteins but act to alter how other cellular genes function after infection. Understanding these viral noncoding RNAs will provide novel targets for the design of therapeutics to combat these cancer-causing viruses.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
5P01CA016038-44
Application #
9512685
Study Section
Special Emphasis Panel (ZCA1)
Project Start
Project End
Budget Start
2018-07-01
Budget End
2019-06-30
Support Year
44
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Yale University
Department
Type
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
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:
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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
Lee, Nara; Yario, Therese A; Gao, Jessica S et al. (2016) EBV noncoding RNA EBER2 interacts with host RNA-binding proteins to regulate viral gene expression. Proc Natl Acad Sci U S A 113:3221-6
DiMaio, Daniel (2016) Thank You, Edward. Merci, Louis. PLoS Pathog 12:e1005320

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