ol Min Jott4 Au Md-Tkn Hnlur Aam Atrhin Analysis of the Hormone-dependent Regulation of a JunD)-Estrogen Receptor Chimcra pl liratiort Februaty Kovemlct 1995/ (Ccc1ol Trecd orm Kevin Ryden Xoy Mary Honald Fhinncys_ Hnc Francisi Wilorda Elamo 9] / Kotuc Far Ccntct Ur AeMcnt unden" nnd junl) elily Jor The moduler ligand-hinding donains of steruid reccp CUM 8lmt, identical, regulation Cenc-Oa Ecncratemolein chimerne trat share high Iore ar rotcis been widely uAcd ~identity (11. '"hcy Ara also sitna tha ligund dependent for #clivily.In acid Finilat nilinie dcirc euttuIO Rpnuchce with c-Fos and Fos-rclated proleins (2). conditionally thcir irteraction Hun) acl Uar Lcncrelcc Rcrice cnlvoxyl (( (OF)- IN at AF'] silcs u ut Jun Cox dimct comnicxcsmd by fusing mt Droldins tficI (OOA-terminal fragmett Jarminn PnCS . Sirco cnGE with Ad rckulia (tc' CutRRi0n LATE4 arid ligund. thntt Juduced (EH) rapidly Juan conuins Uc Trogcn rcctmor amtily ncmticna n TcRuunso extrarciu binding domnin: (FKI cki: Junl)EJe (IKII) nud imnncdiate cark Foir lar Growth siunls tncy nic cinesuice zippcr Hiti iulec Lauci Inic TEun Kxpression IArX Lnce 0 Hamilies is snonE( Ecnt& Eenc exhihit hornon ctivalion aclnttur urO inlcgrul parl of a well-orchcstraled cascadc leading t0 ~onenacn cvoras tcin-]-dirceted trnuecription in transicni exprcSsion MR - ceulat pralilcratior 3nd Tclercnces unortiul Cci fusions, IKK , hae Ont AhteO no nramTICO GhIc the celluler Frowth HEMIAICLAML Darlicinus pruem Uacus dctnil decrmninc tucclaniem nction analyzing thc direrl checls furtoi Eludics hese resunnuro cxperimcnt: IT olfilittureonilion Wte Mrrane Ercatly mroleing 0I FCIIO crnressiol Truwtn wania dcmonstratc that Iromt HEK und Fos iranefccica cclls facilitaled by a wcll characlcrizcd inducible expression syslem. Cur Fos And IKI rendil, arneehntorutinicr omncxtR Onc #pproach for crcating inducible system lucuse : prisingly, the formntion Fos:II heterodimers; plucing te uctivity o protcin under Ihe control ofthc lieatd - IFI #osRInC Fnn] cclimcie troren-inarundcn, sleroid uding dumAI Wanlomo rectuior Ruch Howcvcr, Eel shift ASSAYR clcarly dcmonstrate that I)NA Elucocorlicoid receplor (GJ) AFI sitcs hy Fos:I)ER hctcrodimcrs (hc CstIOgch rereptor (KKA St - IKI hincing crealed wilh proteins hsimi urolems HaVC [icl homodimcra cAtrogon-dopondena.Mor Ovcr, Ihcal- FCI Such os FIA (4), c-Myc (54 the CCAATenhancer-binding un cflectivc inhil GcnCO cetroecn Inc nhi wrotcin (61, Myol) (7), 053 (8L, and c-Fos (9, J0la "f Fosaliuded transuclivudict, this fIfect i6 cach LHced thc #rcscnce the fusiou pratein be' devoid biological Qur Teauils Iudi - HACiunto roicreco ul ceirolem Cheds thc IEI pratein dircct, eXpr"ssed in cellz in thc ahscnco ormono-rovcre- pcliv Mncia M Hgrmnan ible inhibitor of Fos and that estrogen etrols tne con- appropriate wild Vyn (W") functioni was Akcrae iu reclored ncgativc UEI ditione uneilirc 0 dounAn nctivitice harugn in thc prctcncc INA binding thc level thc   possibilit; Fi ? Accordinely Wc explored thc aclivitins WT' end ccll lincs (hat stably cxptoss conely dcrivcrGbrnhlasi could I reculaled tranelationally mamlAtod ruemin Ws thc IFI prulcin exhibit reduced fnlri Iuto tnc Mfuni  (' by fusion with tho FK Uun) chinioric nioloculo R croated quiosccnt of thc cell 0s ni ' Ecruni ema trunculed crclc KK sucuLCO' Han apncndinp' WAA in the Alnent Thic ntad raruren conire thc Jun)-EK (IEI) fusion. Analy-is IEK rerulting suppOrt tha estrogcn-trcated controls_ ncso rcsuil estrocet that for thc Jueinn nrolcn #hout doncnacni crclc hypathesis thnt Al" Bn is imporlant cll uroeros- tranacription from AF] rcporlcr MchAe Constrct examining thc Rion Atid Drovido uniduc AnnTOAOn detailcd analysis of the nicchanism by which esto- addilioml; role of AF'] in thif proccss, IC pcrlorjucd Thc chimera nculale? Vtolci fortur het erodizuet? wilh WT" Fos proteins but docs Jot readily hecruditcrs with WY Jun proicins. Hawevcr, ncither the familics of on oproleins Are tho For Melien LMc' Jun and IEK and W' Jun nor Cna Tulerachons ncucmi Elrone COI[OLCIIL ~ of the dimeric U ausc!iplion "cti- CLCCWE Inctor interactions hctwccn IER and Fos are rCgulaled Vv (stroccn; prateit- (AF'T)" Thc thrcc mucnut no Fut Fetc F corLTasL, IINA binding by HER homodimcrs FERFos complctcly stropen-dependent_ elcrodincn- ccauc thi& novel "clion; thc IFR fusion micchanisu prolein Fwllic Ilcaltl Scrvice Girnml; bupnorlt& JcGative protein that complexcs with Fos cilectvc DuIMMML UaKalatal lunitle Cirmn 32in (e-ng IONA binding: #clivity- Ju addition, #IIc] reversiblt imniituts IS 408u' 4HU:' UaC Eaniceanaran_Amn Cant Moll FSn Glo- cell clones that slably express the IFF prolelo exhibit Myela (Juicolors neinecd Goela puwicalion want by thc payiuchl ailiclt #ert ucitalco cliaigs phasc from Gucr Froxth ATrestcd &lulc uuced CamaCIL arlicle tusl tnciclone hcrony nuirkoFaducelist !ch ( Aaceuroamge rosult thal consislent witn in thc uuscnCC ealracen Ceclion ]734 &lchy Indicntt the hypothesis that AF] Joranal ccll cyclc IIIIiciriatL Felltw Itt Nalional Iuclitulcf Hcalih and nripicng METHII 74-V219 pruprt#SIuI Fcllow _ thc Ancricnn Cancer Socicly: chimeras bclwccn steroid Whilc Mmtecr ticonne 215-726-2889. T tom cotcemandcnccehould WldtetRL 415724.36 C;l, clurocor- TAHedar Mnacd AroA" Actmnr #retm becco'& modilied Kaglcie wc-dium; FHS, Icta Fovine #crun; ]*, thyroid #TI ; wild ty "c; rCipalyuctasc Wiculdrceuigr KK Ualncenecelant rolit oic ecid receptor, HIMKM, Iuk: dreaciol CATchlorennuhonicnl Acoretrenalcrasn horJlOIIC Tccenior_KAK 11502 PUBLICATIONS 0050408
ol Min Jott4 Au Aam Md-Tkn Atrhin Hnlur Analysis of the Hormone-dependent Regulation of a JunD)-Estrogen Receptor Chimcra (Ccc1ol pl liratiort Kovemlct Trecd orm Februaty 1995/ Mary Xoy Francisi Honald Fhinncys_ Hnc Kevin Ryden Kotuc Far Ur AeMcnt Ccntct Wilorda Elamo 9] / The moduler ligand-hinding donains of steruid reccp elily CUM Jor nnd junl) unden" 8lmt, Iore ar Ecncratemolein chimerne identical, regulation Cenc-Oa rotcis been widely uAcd trat share high tha ligund dependent for #clivily.In Finilat nilinie dcirc euttuIO acid Rpnuchce ~identity (11. '"hcy Ara also sitna Uar Lcncrelcc Rcrice conditionally acl Hun) thcir irteraction with c-Fos and Fos-rclated proleins (2). mt Droldins by fusing tficI cnlvoxyl (( (OF)- u ut Jun Cox dimct comnicxcsmd IN at AF'] silcs Jarminn cnGE with (OOA-terminal fragmett Ad rckulia (tc' CutRRi0n LATE4 PnCS . Sirco arid Juan Trogcn rcctmor (EH) thntt conuins Uc ligund. amtily ncmticna rapidly Juduced n TcRuunso extrarciu binding domnin: Junl)EJe (IKII) nud Foir (FKI cki: lar Growth siunls tncy nic cinesuice imnncdiate cark IArX Hiti iulec Lauci zippcr Inic TEun snonE( Ecnt& Kxpression Lnce 0 Eenc Hamilies is exhihit hornon ~onenacn ctivalion aclnttur urO inlcgrul parl of a well-orchcstraled cascadc cvoras leading t0 tcin-]-dirceted trnuecription in transicni exprcSsion MR - ceulat pralilcratior 3nd Tclercnces unortiul Cci GhIc Ont AhteO fusions, IKK , hae no nramTICO pruem HEMIAICLAML Darlicinus the celluler Frowth dctnil decrmninc tucclaniem nction Uacus resunnuro furtoi Eludics analyzing thc direrl checls hese IT olfilittureonilion cxperimcnt: Wte Mrrane mroleing 0I FCIIO crnressiol Truwtn wania dcmonstratc that Ercatly Iromt HEK und Fos iranefccica cclls Cur facilitaled by a wcll characlcrizcd inducible expression syslem. Fos And IKI rendil, arneehntorutinicr omncxtR Onc #pproach for crcating inducible system lucuse : prisingly, the formntion Fos:II heterodimers; plucing te uctivity o protcin under Ihe control ofthc lieatd - #osRInC IFI Fnn] cclimcie troren-inarundcn, uding dumAI sleroid Wanlomo rectuior Ruch Howcvcr, Eel shift ASSAYR clcarly dcmonstrate that I)NA hincing AFI sitcs hy Fos:I)ER hctcrodimcrs IKI Elucocorlicoid receplor (GJ) (hc CstIOgch rereptor (KKA St - homodimcra cAtrogon-dopondena.Mor Ovcr, Ihcal- FCI hsimi urolems HaVC [icl crealed wilh proteins GcnCO cetroecn Inc nhi wrotcin un cflectivc inhil Such os FIA (4), c-Myc (54 the CCAATenhancer-binding "f Fosaliuded transuclivudict, this fIfect i6 (61, Myol) (7), 053 (8L, and c-Fos (9, J0la cach LHced roicreco thc #rcscnce ul ceirolem Qur Teauils Iudi - Cheds the fusiou pratein HACiunto be' devoid biological M thc IEI pratein dircct, ormono-rovcre- pcliv Mncia eXpr"ssed in cellz in thc ahscnco Hgrmnan ible inhibitor of Fos and that estrogen etrols tne con- Akcrae iu appropriate wild Vyn (W") functioni was reclored ditione uneilirc 0 dounAn ncgativc nctivitice UEI in thc prctcncc harugn thc level INA binding Fi ? Accordinely Wc explored thc   possibilit; thc aclivitins WT' end conely dcrivcrGbrnhlasi ccll lincs (hat stably cxptoss mamlAtod ruemin could I reculaled Ws tranelationally thc IFI prulcin exhibit reduced fnlri Iuto tnc Mfuni  (' by fusion with tho FK Uun) chinioric nioloculo R croated of thc cell crclc Han quiosccnt 0s ni ' Ecruni ema apncndinp' trunculed WAA KK sucuLCO' ntad in the Alnent raruren Thic conire rerulting thc Jun)-EK (IEI) fusion. Analy-is IEK estrogcn-trcated controls_ ncso rcsuil suppOrt tha Jueinn nrolcn #hout that estrocet doncnacni for thc hypathesis thnt Al" Bn is imporlant cll crclc uroeros- MchAe tranacription from AF] rcporlcr Constrct Rion Atid Drovido uniduc AnnTOAOn examining thc addilioml; detailcd analysis of the nicchanism by which esto- role of AF'] in thif proccss, nculale? IC Vtolci pcrlorjucd Thc chimera fortur het erodizuet? wilh WT" Fos proteins but docs Jot readily hecruditcrs with WY Jun proicins. Hawevcr, ncither the Melien LMc' Jun and For familics of on oproleins Are tho Cna Tulerachons ncucmi IEK and W' Jun nor Elrone CLCCWE COI[OLCIIL ~ of the dimeric U ausc!iplion Inctor "cti- interactions hctwccn IER and Fos are rCgulaled Vv (stroccn; F prateit- (AF'T)" Thc thrcc mucnut no Fut Fetc corLTasL, IINA binding by HER homodimcrs FERFos elcrodincn- complctcly stropen-dependent_ ccauc thi& novel micchanisu "clion; thc IFR fusion prolein bupnorlt& Fwllic Ilcaltl Scrvice Girnml; UaKalatal lunitle Cirmn cilectvc DuIMMML JcGative protein that complexcs with Fos 32in (e-ng 4HU:' 408u' UaC Eaniceanaran_Amn Cant Moll FSn Glo- #IIc] reversiblt imniituts IS IONA binding: #clivity- Ju addition, Myela (Juicolors neinecd Goela puwicalion cell clones that slably express the IFF prolelo exhibit ailiclt #ert ucitalco want by thc payiuchl cliaigs  uuced CamaCIL Gucr phasc from Froxth ATrestcd &lulc arlicle tusl tnciclone hcrony nuirkoFaducelist !ch ( Aaceuroamge in thc uuscnCC ealracen rosult thal consislent witn Ceclion ]734 &lchy Indicntt METHII the hypothesis that AF] IIIIiciriatL Joranal ccll cyclc Felltw Itt Nalional Iuclitulcf Hcalih and nripicng 74-V219 pruprt#SIuI Fcllow _ thc Ancricnn Cancer Socicly: Whilc Mmtecr ticonne chimeras bclwccn steroid T tom cotcemandcnccehould WldtetRL 215-726-2889. 415724.36 TAHedar Mnacd AroA" Actmnr #retm C;l, clurocor- Wiculdrceuigr KK Ualncenecelant #TI ; wild ty "c; rCipalyuctasc becco'& modilied Kaglcie wc-dium; FHS, Icta Fovine #crun; ]*, thyroid dreaciol CATchlorennuhonicnl Acoretrenalcrasn HIMKM, Iuk: horJlOIIC Tccenior_KAK rolit oic ecid receptor, 11502 PUBLICATIONS 0050408