Stimulation of the human cytomegalovirus IE enhancer/promoter in HL-60 cells by TNFalpha is mediated via induction of NF-kappaB.
TNFalpha enhances the basal activity of the major Immediate Early (IE) enhancer/promoter of human cytomegalovirus (HCMV) in the immature premonocytic HL-60 cell line. The stimulatory effect of TNFalpha is mediated by induction of the transcription factor NF-kappaB, which specifically binds to the 18-bp repetitive sequence motif of the enhancer region. Complex formation could be competed by oligonucleotides representing the 18-bp sequence motif or the prototype NF-kappaB sequence of the immunoglobulin kappa gene. In gel mobility shift assays antisera specific to NF-kappaB p50 and p65 subunits were shown to react with the DNA-protein complex. Addition of the antioxidant PDTC blocked TNFalpha-mediated stimulation in a dose dependent manner. Electrophoretic mobility shift assays indicated that PDTC prevents NF-kappaB induction. Furthermore, it is suggested that protein kinases like PK-C are involved in the TNFalpha signal transduction pathway which leads to the activation of NF-kappaB and its binding to the HCMV IE enhancer in HL-60 cells. Our data are consistent with a role of TNFalpha in reactivation of latent HCMV infection in premonocytic cells.
Prösch
S
S
Institute of Medical Virology, Humboldt University, Charitè Medical School, Berlin, Germany.
Staak
K
K
Stein
J
J
Liebenthal
C
C
Stamminger
T
T
Volk
H D
HD
Krüger
D H
DH
D000956:Antigens, Viral / Q000235:genetics* / Q000378:metabolism; D003587:Cytomegalovirus / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000378:metabolism / Q000821:virology*; D004305:Dose-Response Relationship, Drug; D018922:HL-60 Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects / Q000235:genetics
D000956:Antigens, Viral; D017874:Immediate-Early Proteins; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; C044582:immediate-early proteins, cytomegalovirus