Human T-cell leukemia virus type 1 (HTLV-1) is linked to the genesis of adult T-cell leukemia (ATL), an aggressive CD4+CD25+ malignancy. Tax is an oncoprotein encoded in the pX region of the HTLV-1 genome that regulates viral and cellular gene expression and promotes cell transformation, in part, by constitutive activation of the NF-?B transcription factor. Tax requires NF-?B for the immortalization of primary T cells and HTLV-1 transformed cells are dependent on NF-?B for their survival, thereby providing a strong rationale for studying the mechanisms used by Tax to activate NF-?B. Tax activates the I?B kinase (IKK) and NF-?B persistently by interacting with the IKK regulatory subunit NEMO, however the underlying mechanisms remain poorly understood. Our collective studies reinforce the notion that Tax hijacks the host ubiquitin machinery to activate NF-?B. Tax is polyubiquitinated via Lys63 (K63)-linked polyubiquitin chains and requires the E2 ubiquitin conjugating enzyme Ubc13 for Tax ubiquitination, NEMO binding and NF-?B activation. In our preliminary studies we have conducted ubiquitin proteomics screening which has identified the anti-apoptotic protein Mcl-1 as a downstream target of Tax and the E3 ligase TRAF6. Tax-induced K63-linked polyubiquitination of Mcl-1 endowed Mcl-1 with enhanced stability and prevented its degradation by the chemotherapy drug etoposide. Using a yeast two-hybrid screen we have also identified the E3/E4 enzyme UBE4B as a novel Tax interacting protein that may regulate Tax K63-linked polyubiquitination and NF-?B activation. We have also demonstrated that Tax has usurped the adaptor function of TAX1BP1, a scaffold molecule for the NF-?B inhibitor and ubiquitin-editing enzyme A20, to promote NF-?B signaling. In this competitive renewal, we will continue to establish the mechanisms by which Tax hijacks ubiquitin machinery components to promote persistent NF-?B activation and cell survival.
The specific aims of the proposal are to: (1) determine the mechanisms of Mcl-1 stabilization by Tax; (2) determine the role of the host ubiquitin machinery in Tax-induced NF-?B activation and (3) determine the role of TAX1BP1 in Tax activation of NF-?B and leukemogenesis. These studies will elucidate how a retroviral oncoprotein hijacks host ubiquitination machinery to inhibit cell death and promote cell transformation.

Public Health Relevance

HTLV-1 infection is linked to the genesis of an aggressive malignancy termed adult T-cell leukemia (ATL) after a long latent period. The HTLV-1 genome encodes an oncogenic protein Tax that persistently activates the anti-apoptotic transcription factor NF-?B. This proposal will provide fundamental insight into the mechanisms underlying Tax activation of NF-?B, inhibition of cell death and T-cell transformation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA135362-09
Application #
9264996
Study Section
Molecular Oncogenesis Study Section (MONC)
Program Officer
Read-Connole, Elizabeth Lee
Project Start
2008-07-03
Project End
2019-05-31
Budget Start
2017-06-01
Budget End
2018-05-31
Support Year
9
Fiscal Year
2017
Total Cost
$291,600
Indirect Cost
$111,600
Name
Johns Hopkins University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21205
Mao, Renfang; Yang, Riyun; Chen, Xia et al. (2017) Regnase-1, a rapid response ribonuclease regulating inflammation and stress responses. Cell Mol Immunol 14:412-422
Choi, Young Bong; Shembade, Noula; Parvatiyar, Kislay et al. (2017) TAX1BP1 Restrains Virus-Induced Apoptosis by Facilitating Itch-Mediated Degradation of the Mitochondrial Adaptor MAVS. Mol Cell Biol 37:
Han, Ke-Jun; Foster, Daniel; Harhaj, Edward W et al. (2016) Monoubiquitination of survival motor neuron regulates its cellular localization and Cajal body integrity. Hum Mol Genet 25:1392-405
Bowman, Melissa; Pan, Fan; Harhaj, Edward W (2016) SHARPINing the knowledge of TCR signal control. Nat Immunol 17:221-2
Zhou, Qinjie; Lavorgna, Alfonso; Bowman, Melissa et al. (2015) Aryl Hydrocarbon Receptor Interacting Protein Targets IRF7 to Suppress Antiviral Signaling and the Induction of Type I Interferon. J Biol Chem 290:14729-39
Shembade, Noula; Harhaj, Edward W (2015) Elucidating dynamic protein-protein interactions and ubiquitination in NF-?B signaling pathways. Methods Mol Biol 1280:283-95
Hyun, Jinhee; Ramos, Juan Carlos; Toomey, Ngoc et al. (2015) Oncogenic human T-cell lymphotropic virus type 1 tax suppression of primary innate immune signaling pathways. J Virol 89:4880-93
Jain, Pooja; Lavorgna, Alfonso; Sehgal, Mohit et al. (2015) Myocyte enhancer factor (MEF)-2 plays essential roles in T-cell transformation associated with HTLV-1 infection by stabilizing complex between Tax and CREB. Retrovirology 12:23
Choi, Young Bong; Harhaj, Edward William (2014) Functional implications of mitochondrial reactive oxygen species generated by oncogenic viruses. Front Biol (Beijing) 9:423-436
Choi, Young Bong; Harhaj, Edward William (2014) HTLV-1 tax stabilizes MCL-1 via TRAF6-dependent K63-linked polyubiquitination to promote cell survival and transformation. PLoS Pathog 10:e1004458

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