Inducible expression of IkappaBalpha repressor mutants interferes with NF-kappaB activity and HIV-1 replication in Jurkat T cells

J Biol Chem. 1998 Mar 27;273(13):7431-40. doi: 10.1074/jbc.273.13.7431.


Human immunodeficiency virus (HIV-1) utilizes the NF-kappaB/Rel proteins to regulate transcription through NF-kappaB binding sites in the HIV-1 long terminal repeat (LTR). Normally, NF-kappaB is retained in the cytoplasm by inhibitory IkappaB proteins; after stimulation by multiple activators including viruses, IkappaBalpha is phosphorylated and degraded, resulting in NF-kappaB release. In the present study, we examined the effect of tetracycline-inducible expression of transdominant repressors of IkappaBalpha (TD-IkappaBalpha) on HIV-1 multiplication using stably selected Jurkat T cells. TD-IkappaBalpha was inducibly expressed as early as 3 h after doxycycline addition and dramatically reduced both NF-kappaB DNA binding activity and LTR-directed gene activity. Interestingly, induced TD-IkappaBalpha expression also decreased endogenous IkappaBalpha expression to undetectable levels by 24 h after induction, demonstrating that TD-IkappaBalpha repressed endogenous NF-kappaB-dependent gene transcription. TD-IkappaBalpha expression also sensitized Jurkat cells to tumor necrosis factor-induced apoptosis. De novo HIV-1 infection of Jurkat cells was dramatically altered by TD-IkappaBalpha induction, resulting in inhibition of HIV-1 multiplication, as measured by p24 antigen, reverse transcriptase, and viral RNA. Given the multiple functions of the NF-kappaB/IkappaB pathway, TD-IkappaBalpha expression may interfere with HIV-1 multiplication at several levels: LTR-mediated transcription, Rev-mediated export of viral RNA, inhibition of HIV-1-induced pro-inflammatory cytokines, and increased sensitivity of HIV-1-infected cells to apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anti-Bacterial Agents / pharmacology
  • Apoptosis
  • DNA, Viral / drug effects
  • DNA, Viral / metabolism
  • DNA-Binding Proteins / antagonists & inhibitors*
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics*
  • Down-Regulation / drug effects
  • Doxycycline / pharmacology
  • Gene Expression Regulation, Viral / drug effects
  • HIV Core Protein p24 / metabolism
  • HIV Reverse Transcriptase / metabolism
  • HIV-1 / genetics
  • HIV-1 / physiology*
  • Humans
  • I-kappa B Proteins*
  • Jurkat Cells
  • Molecular Sequence Data
  • NF-KappaB Inhibitor alpha
  • NF-kappa B / antagonists & inhibitors*
  • RNA, Viral / drug effects
  • RNA, Viral / metabolism
  • Repressor Proteins / pharmacology*
  • T-Lymphocytes / virology
  • Virus Replication / drug effects*


  • Anti-Bacterial Agents
  • DNA, Viral
  • DNA-Binding Proteins
  • HIV Core Protein p24
  • I-kappa B Proteins
  • NF-kappa B
  • NFKBIA protein, human
  • RNA, Viral
  • Repressor Proteins
  • NF-KappaB Inhibitor alpha
  • HIV Reverse Transcriptase
  • Doxycycline