The roles of non-coding RNAs in lymphoid cells harboring three transforming herpesviruses are being investigated. Epstein-Barr virus (EBV) infects and transforms humanB 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 human and monkey T lymphocytes in culture, generating a mature;activated phenotype. Kaposi's sarcoma-associated herpesvirus (KSHV) afflicts immunocompromised individuals and persists in a latent form until lytic activation. The two EBVencoded EBERs, as well as >23 newly identified microRNAs, and the seven HVS-encoded HSURs are expressed in virally transformed cells. Upon induction, KSHV produces PAN, a capped, polyadenylated RNA that does not leave the nucleus. These viral RNAs are all small (from 21 nts to 1.1 kb), abundant, conserved, and associate with host proteins to form RNPs. Studying their functions has,already contributed important insights into host cell pathways perturbed during viral transformation or lytic growth. Proposed aims will test the hypotheses that EBERs, HSURs and PAN manipulate cellular metabolism by sequestering host proteins or microRNAs, or that the resulting RNPs perform a critical viral function. We recently found that HSUR RNPs bind certain host microRNAs;interference with their regulatory roles in transformed T cells will be examined. The possibility that EBERs likewise bind host microRNAs will be tested. Our discovery that microRNAs in quiescent cells activate rather than repress translation will be exploited to investigate the EBV-encoded microRNAs since many transformed B cells exist in a quiescent state in the body. We showed that PAN RNA accumulates to very high nuclear levels in KSHV-reactivated cells because an element (the ENE) near its 3'end engages the polyA tail to prevent RNA decay. Structural studies of the ENE +/- oligoA, as well as complexed with polyA binding protein (PABPC1), will provide mechanistic insights. Our observation that PAN RNA is involved in relocalization of host PABPC1 to the nucleus, suggesting collaboration with the KSHV SOX protein in host mRNA shut off, will be investigated

Public Health Relevance

We are studying how viruses 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 genes function after infection. Understanding these viral non-coding 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 #
5P01CA016038-40
Application #
8476993
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
$162,241
Indirect Cost
$63,114
Name
Yale University
Department
Type
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Guo, Yang Eric; Steitz, Joan A (2014) 3'-Biotin-tagged microRNA-27 does not associate with Argonaute proteins in cells. RNA 20:985-8
Goodwin, Edward C; Motamedi, Nasim; Lipovsky, Alex et al. (2014) Expression of DNAJB12 or DNAJB14 causes coordinate invasion of the nucleus by membranes associated with a novel nuclear pore structure. PLoS One 9:e94322
Cech, Thomas R; Steitz, Joan A (2014) The noncoding RNA revolution-trashing old rules to forge new ones. Cell 157:77-94
Dimaio, Daniel (2014) Is virology dead? MBio 5:e01003-14
Guo, Yang Eric; Riley, Kasandra J; Iwasaki, Akiko et al. (2014) Alternative capture of noncoding RNAs or protein-coding genes by herpesviruses to alter host T cell function. Mol Cell 54:67-79
Carney, Daniel W; Nelson, Christian D S; Ferris, Bennett D et al. (2014) Structural optimization of a retrograde trafficking inhibitor that protects cells from infections by human polyoma- and papillomaviruses. Bioorg Med Chem 22:4836-47
Zhang, Wei; Kazakov, Teymur; Popa, Andreea et al. (2014) Vesicular trafficking of incoming human papillomavirus 16 to the Golgi apparatus and endoplasmic reticulum requires ?-secretase activity. MBio 5:e01777-14
Xie, Mingyi; Steitz, Joan A (2014) Versatile microRNA biogenesis in animals and their viruses. RNA Biol 11:673-81
Park, Richard; El-Guindy, Ayman; Heston, Lee et al. (2014) Nuclear translocation and regulation of intranuclear distribution of cytoplasmic poly(A)-binding protein are distinct processes mediated by two Epstein Barr virus proteins. PLoS One 9:e92593
Guo, Yang Eric; Steitz, Joan A (2014) Virus meets host microRNA: the destroyer, the booster, the hijacker. Mol Cell Biol 34:3780-7

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