Amylloi hei.i-protem (AU) is Ihe major proleineaceous component of the amyloid deposits that accumulate extracelluiarly in the brain of Al/heirncr's disease (AD). A is generated proieolyik-ally from a larger amyloid precursor proteins (BAPP). The apparent over expression of llie UAl'P gene in certain areas of AD brains indicate that abnormalities in gene regulation might he .in important I'arior in AD. Study of the structure and function of the IJAPi1 gene-promoter is an important urea ot" research. Here I report that an upstream regulatory element <URE) located between -22H t" -22fi5 of the human BAPP promoter may interact Mlh a novel proicm (s), U REP. as determined hy the eleclrophorelic mobility shift assay (EMSA). To determine if UREP is related to an already characleriJ.ed transcription fat-tor, EMSA wjs peitnimed iixiiig various -pecil'ic competitors. The labeled URE prohe could interact wiih a disinui mide.ii- factor which was no l cumpcMed hy l lie oligonncleolide.s specific for ditk-rem transcription tacu.rs. AI'l. M'2. Al'3. Ocl-l, NF-1, NF-kB. ORE. and alternatively Hie speeihc pmieiti h.nul M deeded wiili tilher the labeled NF-kB or NF-I probe could not he competed out with excess unlalvlcd URE. To cleleiniuie if such a UREP hand could he detected ID human brain li-siie samples, EMSA from Hie nuclear extracts of the human hrain was pcTloiiiK-d. A disiinu URE specific nuclear factor w.is detected in ditferen! regions of the brain .is well. To (leieiiiimc ilie si/e of ilK- proiem(s) that were specifically bound in Ihe DNApi.ilun uiniplexcs. Soiilhwestein blotting was peiformed. Using Ihe URE probe. Iwo protein bands ol - M kHa .nul l l f. kDa wvio detected m PC12 nuclear extracts. A pCATP/URE reeombin.ini pl.iMind w.is made hy svihcloiung URE in an enliancerless promoter vector pCATl'. I'Ci: cells ih.n were traiisfccted wiih pCATP/ URE displayed 34-fold mon- promoter .Ktivny Hum i-olls thai were iraiislVcR'd with pCATl1. These results suggest that the URE seiiueiifc displayed .in enhjiifer-like activity, lh.it the UREP factor interacting with the URE is no! related to the known transcription factors tested, and that Die protein is expressed in various .ell \\,vs ,iid ,1,Ik-rent rmnv of Ihe huin.m bum. Supported b> a N1H grant.
|Original language||English (US)|
|State||Published - Dec 1 1997|
ASJC Scopus subject areas
- Molecular Biology