Promoter for neuropeptide FF and use thereof for therapy and diagnostics

DETAILED DESCRIPTION OF THE INVENTION We report here the cloning of the 5'-flanking region of the NPFF gene from mouse, rat and human.
Totally 9.
8 kb, 1.
5 kb and 1.
3 kb of the NPFF promoter from mouse, rat and human was cloned and sequenced.
Comparisons between the promoter region from all species showed a high sequence similarity.
Such a high sequence homology and well-conserved structure could indicate an important physiological function and a need for a similar, tightly regulated transcription of the gene.
We have previously shown that the NPFF gene is expressed in specific regions in the brain and in the spinal cord and is induced upon inflammatory stimulus (Vilim et al.
, 1999).
In agreement with this several inflammation related transcription factor consensus sites were found (FIG.
1B), amongst them e.
g.
NF.
kappa.
B, which is considered as an immediate early mediator of immune and inflammatory responses (Lenardo & Baltimore, 1989).
Also several consensus sites for the nuclear factor of activated T-cells (NFAT) were found (for review see Kel et al.
, 1999).
Transcription factors belonging to this growing family play key roles in the regulation of cytokine and other genes during immune response.
An interesting finding was also the consensus site in mouse and rat for heat shock factor 1 (HSF1).
HSF1 is activated in cells exposed to elevated temperatures and other environmental stress conditions (Sarge et al.
, 1993).
We also focused on the potential involvement of AP1, STAT1 and CREB.
By using transiently transfected cells with deletional series of mouse NPFF promoter fused with firefly luciferase cDNA we could show that the basal activity of the promoter changes with increasing promoter size (FIG.
2B).
This indicates that the promoter is tightly regulated is affected by several transcriptional factors and apparently has a complex transcriptional control.
To study the possible involvement of some transcription factors our attempt was to a) activate transcription factors by protein kinase C (PKC) b) activate NFATp by PKC and Ca