TY - JOUR

T1 - Oligonucleotide fishing for STAT6: cross-talk between IL-4 and chemokines.

AU - Eriksen, K W

AU - Nielsen, M

AU - Kaltoft, K

AU - Svejgaard, A

AU - Nissen, Mogens Holst

AU - Röpke, C

AU - Ødum, N

N1 - Keywords: Animals; Base Sequence; Binding Sites; CD4-Positive T-Lymphocytes; Cell Line; Chemokines; Cytokines; DNA; Humans; Interleukin-4; Mice; Molecular Sequence Data; Oligonucleotide Probes; Phosphorylation; STAT6 Transcription Factor; Trans-Activators; Tyrosine

PY - 2001

Y1 - 2001

N2 - Signal transducer and activator of transcription 6 (STAT6) is essential for the biological activities of interleukin-4 (IL-4) and the development of allergic responses in mice. Here we report on a sensitive and specific assay for STAT6 activation in response to IL-4. We took advantage of double-stranded oligonucleotide probes containing a STAT6-binding gene-sequence from the promotor of the immunoglobulin heavy chain germline epsilon transcript to study the IL-4-induced DNA binding of STAT6. Using these probes, we show that repeated adjacent STAT6-binding sites result in enhanced STAT6-DNA binding. Moreover, the distance between the binding sites is critical for STAT-DNA binding, i.e. STAT6 binding is decreased at distances above 20 nucleotides between neighbouring binding sites. Using this assay to study cross-talk between IL-4 and chemokines, we provide evidence that MIP-1beta and MIG inhibit IL-4-induced STAT6 activation, whereas other chemokines and cytokines do not. In conclusion, our data show that oligonucleotide fishing is a supplementary tool for studying cytokine cross-talk at a genomic level.

AB - Signal transducer and activator of transcription 6 (STAT6) is essential for the biological activities of interleukin-4 (IL-4) and the development of allergic responses in mice. Here we report on a sensitive and specific assay for STAT6 activation in response to IL-4. We took advantage of double-stranded oligonucleotide probes containing a STAT6-binding gene-sequence from the promotor of the immunoglobulin heavy chain germline epsilon transcript to study the IL-4-induced DNA binding of STAT6. Using these probes, we show that repeated adjacent STAT6-binding sites result in enhanced STAT6-DNA binding. Moreover, the distance between the binding sites is critical for STAT-DNA binding, i.e. STAT6 binding is decreased at distances above 20 nucleotides between neighbouring binding sites. Using this assay to study cross-talk between IL-4 and chemokines, we provide evidence that MIP-1beta and MIG inhibit IL-4-induced STAT6 activation, whereas other chemokines and cytokines do not. In conclusion, our data show that oligonucleotide fishing is a supplementary tool for studying cytokine cross-talk at a genomic level.

M3 - Journal article

C2 - 11872954

VL - 18

SP - 233

EP - 241

JO - Experimental and Clinical Immunogenetics

JF - Experimental and Clinical Immunogenetics

SN - 0254-9670

IS - 4

ER -