Inhibition of NF-kappa B by a cell permeable form of I kappa B alpha induces apoptosis in eosinophils.
An 11 amino acid HIV-TAT peptide can deliver target proteins into a variety of cells in a receptor-independent manner. To generate a highly specific inhibitor of the transcription factor NF-kappa B, we have fused the TAT-peptide to a version of I kappa B alpha that is resistant to signal-induced degradation. TAT-I kappa B alpha(S32A, S36A) inhibited NF-kappa B-dependent transcription in HeLa and A549 cells by retaining NF-kappa B p65 in the cytoplasm. Introduction of TAT-I kappa B alpha(S32A, S36A) into human eosinophils inhibited the nuclear translocation of NF-kappa B and induced apoptosis. Thus, continuous NF-kappa B-dependent transcription is important for eosinophil survival. While eosinophils are normally refractive to standard methods of gene delivery, the ability of TAT fusion proteins to be taken up by these cells should enable a detailed molecular analysis of survival pathways in these cells.
Fujihara
Satoko
S
Department of Medicine, Centre for Inflammation Research, University of Edinburgh Medical School, Teviot Place, Edinburgh EH8 9AG, UK. s.fujihara@dundee.ac.uk
Jaffray
Ellis
E
Farrow
Stuart N
SN
Rossi
Adriano G
AG
Haslett
Christopher
C
Hay
Ronald T
RT
D017209:Apoptosis / Q000502:physiology*; D002467:Cell Nucleus / Q000378:metabolism; D004804:Eosinophils / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling
D005456:Fluorescent Dyes; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D000072000:NF-KappaB Inhibitor alpha; D016650:Fluorescein-5-isothiocyanate