Low intracellular zinc impairs the translocation of activated NF-kappa B to the nuclei in human neuroblastoma IMR-32 cells.
In the current work, we studied how variations in extracellular zinc concentrations modulate different steps involved in nuclear factor kappaB (NF-kappaB) activation in human neuroblastoma IMR-32 cells. Cells were incubated in media containing varying concentrations of zinc (1.5, 5, 15, and 50 microm). Within 3 h, the intracellular zinc content was lower in cells exposed to 1.5 and 5 microm, compared with the other groups. Low intracellular zinc concentrations were associated with the activation of NF-kappaB, based on high levels of IkappaBalpha phosphorylation, low IkappaBalpha concentrations, and high NF-kappaB binding activity in total cell fractions. However, the active dimer accumulated in the cytosol, as shown by a low ratio of nuclear/cytosolic NF-kappaB binding activity. This altered nuclear translocation was accompanied by a decreased transactivation of an endogenous NF-kappaB-driven gene (ikba) and of a reporter gene (pNF-kappaB-luc). In cells with low intracellular zinc concentrations, a low rate of in vitro tubulin polymerization was measured compared with the other groups. We conclude that low intracellular zinc concentrations induce tubulin depolymerization, which may be one signal for NF-kappaB activation. However, NF-kappaB nuclear translocation is impaired, which inhibits the transactivation of NF-kappaB-driven genes. This could affect cell survival, and be an important factor in certain zinc deficiency-associated pathologies.
Mackenzie
Gerardo G
GG
Departamento de Química Biológica, Instituto de Química y Físicoquímica Biológicas, Facultad de Farmacia y Bioquimica, Universidad de Buenos Aires, C1113AAD Buenos Aires, Argentina.
Zago
M Paola
MP
Keen
Carl L
CL
Oteiza
Patricia I
PI
D021581:Active Transport, Cell Nucleus; D017209:Apoptosis; D003546:Cysteine Endopeptidases / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics; D006801:Humans; D020672:I-kappa B Proteins; D009097:Multienzyme Complexes / Q000502:physiology; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism*; D009447:Neuroblastoma / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D014404:Tubulin / Q000378:metabolism; D015032:Zinc / Q000172:deficiency / Q000502:physiology*
D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D009097:Multienzyme Complexes; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D014404:Tubulin; D000072000:NF-KappaB Inhibitor alpha; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D015032:Zinc