Abstract
To determine critical host factors involved in HIV-1 replication, a dominant effector genetics approach was developed to reveal signaling pathways on which HIV-1 depends for replication. A large library of short peptide aptamers was expressed via retroviral delivery in T cells. Peptides that interfered with T cell activation-dependent processes that might support HIV-1 replication were identified. One of the selected peptides altered signaling, lead to a difference in T cell activation status, and inhibited HIV-1 replication. The target of the peptide was JAB1/CSN5, a component of the signalosome complex. JAB1 expression overcame the inhibition of HIV-1 replication in the presence of peptide and also promoted HIV-1 replication in activated primary CD4(+) T cells. This peptide blocked physiological release of JAB1 from the accessory T cell surface protein LFA-1, downstream AP-1 dependent events, NFAT activation, and HIV-1 replication. Thus, genetic selection for intracellular aptamer inhibitors of host cell processes proximal to signals at the immunological synapse of T cells can define unique mechanisms important to HIV-1 replication.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Aptamers, Peptide / chemistry
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Aptamers, Peptide / pharmacology
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CD4-Positive T-Lymphocytes / drug effects
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CD4-Positive T-Lymphocytes / enzymology
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CD4-Positive T-Lymphocytes / pathology
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CD4-Positive T-Lymphocytes / virology
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COP9 Signalosome Complex
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HIV-1 / drug effects
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HIV-1 / genetics
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HIV-1 / physiology*
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Humans
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Intracellular Signaling Peptides and Proteins / metabolism*
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Intracellular Space / drug effects
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Intracellular Space / metabolism
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JNK Mitogen-Activated Protein Kinases / metabolism
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Jurkat Cells
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Lymphocyte Activation / drug effects
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Lymphocyte Function-Associated Antigen-1 / metabolism*
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Models, Immunological
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Molecular Sequence Data
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Multiprotein Complexes / metabolism*
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NFATC Transcription Factors / metabolism
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Peptide Hydrolases / metabolism*
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Peptide Library
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Protein Binding / drug effects
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Protein Transport / drug effects
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Retroviridae / genetics
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Signal Transduction* / drug effects
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T-Lymphocytes / drug effects
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T-Lymphocytes / enzymology
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T-Lymphocytes / pathology
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T-Lymphocytes / virology*
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Transcription Factor AP-1 / metabolism
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Transcription, Genetic / drug effects
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Virus Replication / drug effects
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Virus Replication / physiology*
Substances
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Aptamers, Peptide
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Intracellular Signaling Peptides and Proteins
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Lymphocyte Function-Associated Antigen-1
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Multiprotein Complexes
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NFATC Transcription Factors
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Peptide Library
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Transcription Factor AP-1
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JNK Mitogen-Activated Protein Kinases
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Peptide Hydrolases
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COPS5 protein, human
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COP9 Signalosome Complex