Intranuclear targeted delivery of functional NF-kappaB by 70 kDa heat shock protein.
The 70 kDa heat shock protein (Hsp70) is a highly conserved, ubiquitous protein involved in chaperoning proteins to various cellular organelles. Here we show that when added exogenously to cells, Hsp70 is readily imported into both cytoplasmic and nuclear compartments in a cell-type-specific fashion. We exploited this ability of Hsp70 to deliver NF-kappaB, a key transcriptional regulator of inflammatory responses. We demonstrate that a fusion protein composed of a C-terminal Hsp70 peptide and the p50 subunit of NF-kappaB was directed into the nucleus of cells, could bind DNA specifically, and activated Igkappa expression and TNFalpha production. We therefore propose that Hsp70 can be used as a vehicle for intracytoplasmic and intranuclear delivery of proteins or DNA to modulate gene expression and thereby control immune responses.
Fujihara
S M
SM
Bristol-Myers Squibb Pharmaceutical Research Institute, PO Box 4000, Princeton, NJ 08543, USA.
Nadler
S G
SG
D001402:B-Lymphocytes / Q000378:metabolism; D001483:Base Sequence; D001693:Biological Transport, Active; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D009000:Monocytes / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D007145:Immunoglobulin kappa-Chains; D016328:NF-kappa B; D011947:Receptors, Antigen, B-Cell; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D004247:DNA