Dynamic analysis of STAT6 signalling in living cells.
Functional activity of N- and C-terminal fluorescent fusion proteins between STAT6 and EGFP was demonstrated through IL-4-dependent transcriptional activation and nuclear translocation. The N-terminal (EGFP-STAT6) fusion protein appeared to be more active than the C-terminal fusion. In HEK-293 cells both fusion proteins formed fluorescent nuclear foci following IL-4 stimulation, but in HeLa cells nuclear accumulation was homogeneous. Stimulation of the NF-kappaB pathway through TNFalpha treatment, or expression of p65-EGFP fusion protein, repressed both basal STAT6-dependent transcriptional activity and the extent of activation in response to IL-4. This indicates a novel mechanism of inhibition of STAT6 signalling by NF-kappaB activation.
Nelson
Glyn
G
School of Biological Sciences, Life Sciences Building, University of Liverpool, Crown St., Liverpool L69 7ZB, UK.
Wilde
Geraint J C
GJ
Spiller
David G
DG
Sullivan
Elaine
E
Unitt
John F
JF
White
Michael R H
MR
D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D016328:NF-kappa B / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D050800:STAT6 Transcription Factor; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D051996:Transcription Factor RelA; D015533:Transcriptional Activation
D008164:Luminescent Proteins; D016328:NF-kappa B; D011993:Recombinant Fusion Proteins; D050800:STAT6 Transcription Factor; C494101:STAT6 protein, human; D015534:Trans-Activators; D051996:Transcription Factor RelA; D049452:Green Fluorescent Proteins; D015847:Interleukin-4