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Regulation of NFk B , AP-1 , NFAT , and STAT 1 Nuclear Import in T Lymphocytes by Noninvasive Delivery of Peptide Carrying the Nuclear Localization Sequence of NFk B p 501

Activation of T lymphocytes by Ags or cytokines results in translocation of the transcription factors NFkB, AP-1, NFAT, and STAT from the cytoplasm into the nucleus. The first step in the nuclear import process is recognition of a nuclear localization sequence (NLS) within the karyophilic protein by a cytoplasmic receptor such as the importin (karyopherin)a subunit. The NLSs of NF-kB, AP-1, and NFAT differ and the NLS of STAT1 has not yet been identified. Herein we demonstrate that the inducible nuclear import of NF-kB, AP-1, NFAT, and STAT1 in Jurkat T lymphocytes is significantly inhibited by a cell-permeable peptide carrying the NLS of the NF-kB p50 subunit. NLS peptide-mediated disruption of the nuclear import of these transcription factors results in inhibition of I kBa and IL-2 gene expression, processes dependent on NFkB or the combination of NF-kB, AP-1, and NFAT. Further, we show that inhibitory NLS peptide interacts in vitro with a cytoplasmic NLS receptor complex comprised of the Rch1/importin (karyopherin)b heterodimer expressed in Jurkat T cells. Taken together, these data indicate that the inducible nuclear import of NF-kB, AP-1, NFAT, and STAT1 in Jurkat T cells can be regulated by NLS peptide delivered noninvasively to the cytoplasm of Jurkat T cells to target members of the importin (karyopherin)-ab NLS receptor complex. The Journal of Immunology,1998, 161: 6084–6092.