Major stress protein Hsp70 interacts with NF-kB regulatory complex in human T-lymphoma cells.
Polypeptides belonging to the Hsp70 major stress protein family and to the NF-kB/Rel multi-functional regulatory complex are known to be involved in cellular defense mechanisms. It was suggested that both systems may interact in cells that respond to injuring stimuli. To check this, Molt4 human lymphoma cells were heated at 43 degrees C for 15 min and, after a 6 h post-shock recovery period, the cells were activated with phorbol ester or bacterial lipopolysaccharide. It was found that mild heat shock caused a substantial increase of the intracellular Hsp70 content with the concomitant suppression of NF-kB complexes, though the latter was properly activated in non-stressed cells. After a 24 h period of being inactive the complex fully recovered its activity and p65 and c-Rel subunits migrated to the nucleus. This new active period lasted even longer than that in non-heated control cells. As this suggested the existence of a Hsp70-related mechanism of NF-kB/Rel complex retention in cytoplasm, we carried out immunoprecipitation with the use of anti-Hsp70 and anti-Rel antibodies. All three Rel family members p65, c-Rel, p50, but not their precursors and IkB alpha inhibitory protein were shown to co-precipitate with the stress protein and anti-Hsp70 antibodies from both heated and non-heated cells. We conclude that the Hsp70 stress protein may confer a new mechanism of NF-kB regulation in cells affected by elevated temperature or other factors related to the cellular response to stress.
Guzhova
I V
IV
Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos, Brazil. ira@uenf.br
Darieva
Z A
ZA
Melo
A R
AR
Margulis
B A
BA
D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D016399:Lymphoma, T-Cell; D016328:NF-kappa B / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D020569:Proto-Oncogene Proteins c-rel; D013312:Stress, Physiological / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism
D018840:HSP70 Heat-Shock Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; D010703:Phorbol Esters; D011518:Proto-Oncogene Proteins; D020569:Proto-Oncogene Proteins c-rel; D014157:Transcription Factors