Abstract
Large T antigen (TAg) of the human polyomavirus JC virus (JCV) possesses DNA binding and helicase activities, which, together with various cellular proteins, are required for replication of the viral genome. We now show that JCV-infected cells expressing TAg accumulate in the G(2) phase of the cell cycle as a result of the activation of ATM- and ATR-mediated G(2) checkpoint pathways. Transient transfection of cells with a TAg expression vector also induced G(2) checkpoint signaling and G(2) arrest. Analysis of TAg mutants with different subnuclear localizations suggested that the association of TAg with cellular DNA contributes to the induction of G(2) arrest. Abrogation of G(2) arrest by inhibition of ATM and ATR, Chk1, and Wee1 suppressed JCV genome replication. In addition, abrogation of the G(2)-M transition by Cdc2 depletion disabled Wee1 depletion-induced suppression of JCV genome replication, suggesting that JCV replication is facilitated by G(2) arrest resulting from G(2) checkpoint signaling. Moreover, inhibition of ATM and ATR by caffeine suppressed JCV production. The observation that oligodendrocytes productively infected with JCV in vivo also undergo G(2) arrest suggests that G(2) checkpoint inhibitors such as caffeine are potential therapeutic agents for JCV infection.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Antigens, Viral, Tumor / genetics
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Antigens, Viral, Tumor / metabolism*
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Ataxia Telangiectasia Mutated Proteins
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CDC2 Protein Kinase
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Cell Cycle Proteins / antagonists & inhibitors
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Cell Cycle Proteins / genetics
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Cell Cycle Proteins / metabolism*
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Cell Line, Tumor
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Checkpoint Kinase 1
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Cyclin B / genetics
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Cyclin B / metabolism
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Cyclin-Dependent Kinases
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DNA-Binding Proteins / antagonists & inhibitors
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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G2 Phase*
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Genome, Viral / physiology
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Humans
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JC Virus / physiology*
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Mutation
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Nuclear Proteins
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Oligodendroglia / metabolism
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Oligodendroglia / virology
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Polyomavirus Infections / drug therapy
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Polyomavirus Infections / genetics
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Polyomavirus Infections / metabolism
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Protein Kinases / genetics
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Protein Kinases / metabolism
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Protein Serine-Threonine Kinases / antagonists & inhibitors
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Protein-Tyrosine Kinases
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Signal Transduction*
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Tumor Suppressor Proteins / antagonists & inhibitors
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Tumor Suppressor Proteins / genetics
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Tumor Suppressor Proteins / metabolism*
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Tumor Virus Infections / drug therapy
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Tumor Virus Infections / genetics
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Tumor Virus Infections / metabolism
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Virus Replication / physiology*
Substances
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Antigens, Viral, Tumor
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Cell Cycle Proteins
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Cyclin B
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DNA-Binding Proteins
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Nuclear Proteins
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Tumor Suppressor Proteins
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Protein Kinases
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Protein-Tyrosine Kinases
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WEE1 protein, human
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ATM protein, human
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ATR protein, human
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Ataxia Telangiectasia Mutated Proteins
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CHEK1 protein, human
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Checkpoint Kinase 1
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Protein Serine-Threonine Kinases
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CDC2 Protein Kinase
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CDK1 protein, human
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Cyclin-Dependent Kinases