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B, AP-1, NFAT, and STAT1 Nuclear Import in T Lymphocytes by Noninvasive Delivery of Peptide Carrying the Nuclear Localization Sequence of NF-B p501
Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN 37232
Activation of T lymphocytes by Ags or cytokines results in
translocation of the transcription factors NF-
B, 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)-
subunit. The NLSs of NF-B, 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-B, AP-1, NFAT,
and STAT1 in Jurkat T lymphocytes is significantly inhibited by a
cell-permeable peptide carrying the NLS of the NF-B p50 subunit. NLS
peptide-mediated disruption of the nuclear import of these
transcription factors results in inhibition of IB and IL-2 gene
expression, processes dependent on NF-B or the combination of
NF-B, 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)-ß heterodimer expressed in Jurkat T
cells. Taken together, these data indicate that the inducible nuclear
import of NF-B, 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)-ß
NLS receptor complex.
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