JC virus (JCV) is the etiologic agent of the neurodegenerative disease, progressive multifocal leukoencephalopathy (PML). JCV is normally latent in non-immunocompromised people, but it is activated in brains of individuals with AIDS. Because of the high incidence of PML in AIDS, we have hypothesized that activation of JCV in glial cells of the brain is influenced by HIV-1 infection. We have found that JCV late gene transcription is stimulated by the HIV-1 Tat protein through action at sequence elements bound by the cellular protein, Pur about. Tat and Pur0 about form a highly specific complex, and this complex acts to stimulate transcription at both the HW-1 TAR RNA element and the JCV late promoter. The complex also interacts with T-antigen to enhance JCV DNA replication. Purc about is a frequent partner of cyclin/CDK complexes, as is another Tat-binding protein, cyclin T1. This proposal aims to continue to exploit the expertise of two independent laboratories to coordinate studies on HIV-1, JCV and cellular proteins, including Pur0 about and cyclin T1. We shall take advantage of rapid autopsy procedures at the Manhattan HIV Brain Bank to perform immunogold electron microscopy on PML brain samples to colocalize Tat with these cellular proteins. We shall elucidate the dynamics of the interactions between Tat, T-antigen and Puro about during the course of JCV infection of oligodendrocytes and determine whether transcriptional activation involves activity of cyclin T1/CDK9. We shall characterize the involvement of Purc about or cyclin T1/CDK9 in Tat activation of transcription at TAR(+) or TAR(-) HIV-1 LTR promoters. Use of Puro about point mutants in AA V vectors will help dissect the contributions of different molecular unctional groups to HIV-1 replication in microglial/monocytic cells. We shall pursue our finding that exogenous Tat at low concentrations is incorporated by KG-1 oligodendrocytes and stimulates DNA replication initiated at the JCV origin. We shall determine the mechanism by which Tat enhances replication initiated at the JCV origin in human oligodendrocytes both in vivo and using a new in vitro system. Results will help elucidate pathways of activation of HIV-1 and JCV in the brain and will help target particular molecular interactions for therapy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS035000-06A2
Application #
6450231
Study Section
AIDS and Related Research 8 (AARR)
Program Officer
Nunn, Michael
Project Start
1996-02-01
Project End
2007-01-31
Budget Start
2002-02-01
Budget End
2003-01-31
Support Year
6
Fiscal Year
2002
Total Cost
$371,691
Indirect Cost
Name
Mount Sinai School of Medicine
Department
Pathology
Type
Schools of Medicine
DUNS #
114400633
City
New York
State
NY
Country
United States
Zip Code
10029
Daniel, Dianne C; Johnson, Edward M (2018) PURA, the gene encoding Pur-alpha, member of an ancient nucleic acid-binding protein family with mammalian neurological functions. Gene 643:133-143
Johnson, Edward M; Wortman, Margaret J; Dagdanova, Ayuna V et al. (2013) Polyomavirus JC in the context of immunosuppression: a series of adaptive, DNA replication-driven recombination events in the development of progressive multifocal leukoencephalopathy. Clin Dev Immunol 2013:197807
Wright, Clayton A; Nance, Jonas A; Johnson, Edward M (2013) Effects of Tat proteins and Tat mutants of different human immunodeficiency virus type 1 clades on glial JC virus early and late gene transcription. J Gen Virol 94:514-23
Darbinyan, Armine; Kaminski, Rafal; White, Martyn K et al. (2013) Polyomavirus JC infection inhibits differentiation of oligodendrocyte progenitor cells. J Neurosci Res 91:116-27
Ferenczy, Michael W; Marshall, Leslie J; Nelson, Christian D S et al. (2012) Molecular biology, epidemiology, and pathogenesis of progressive multifocal leukoencephalopathy, the JC virus-induced demyelinating disease of the human brain. Clin Microbiol Rev 25:471-506
Noch, Evan; Sariyer, Ilker Kudret; Gordon, Jennifer et al. (2012) JC virus T-antigen regulates glucose metabolic pathways in brain tumor cells. PLoS One 7:e35054
Merabova, Nana; Kaminski, Rafal; Krynska, Barbara et al. (2012) JCV agnoprotein-induced reduction in CXCL5/LIX secretion by oligodendrocytes is associated with activation of apoptotic signaling in neurons. J Cell Physiol 227:3119-27
Tavazzi, E; Ferrante, P; Khalili, K (2011) Progressive multifocal leukoencephalopathy: an unexpected complication of modern therapeutic monoclonal antibody therapies. Clin Microbiol Infect 17:1776-80
Wortman, Margaret J; Hanson, Laura K; Martinez-Sobrido, Luis et al. (2010) Regulation of PURA gene transcription by three promoters generating distinctly spliced 5-prime leaders: a novel means of fine control over tissue specificity and viral signals. BMC Mol Biol 11:81
Johnson, Edward M (2010) Structural evaluation of new human polyomaviruses provides clues to pathobiology. Trends Microbiol 18:215-23

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