Abstract
Uncoupling protein 3 (UCP3) is mainly expressed in muscle. It plays an important role in muscle, but less research on the regulation of cattle UCP3 has been performed. In order to elucidate whether cattle UCP3 can be regulated by muscle-related factors, deletion of cattle UCP3 promoter was amplified and cloned into pGL3-basic, pGL3-promoter and PEGFP-N3 vector, respectively, then transfected into C2C12 myoblasts cells and UCP3 promoter activity was measured using the dual-Luciferase reporter assay system. The results showed that there is some negative-regulatory element from −620 to −433 bp, and there is some positive-regulatory element between −433 and −385 bp. The fragment (1.08 kb) of UCP3 promoter was cotransfected with muscle-related transcription factor myogenic regulatory factors (MRFs) and myocyte-specific enhancer factor 2A (MEF2A). We found that UCP3 promoter could be upregulated by Myf5, Myf6 and MyoD and downregulated by MyoG and MEF2A.
Highlights
Uncoupling protein 3 (UCP3) is a member of the family of uncoupling proteins (UCPs), which are members of the mitochondrial anion carrier family
The results showed that Myf5, Myf6 and MyoD contributed to the up-regulation of the UCP3 expression in C2C12 myoblasts, which was consistent with the present results that MyoD was required to activate the promoter of human UCP3 [5]
The results showed that UCP3 promoter led to tissue-specific expression in the muscle
Summary
Uncoupling protein 3 (UCP3) is a member of the family of uncoupling proteins (UCPs), which are members of the mitochondrial anion carrier family. Uncoupling protein-3 (UCP3) gene is primarily expressed in skeletal muscle and up-regulated by fatty acids [5]. MyoD controls UCP3 promoter activity through a noncanonical E box site located close to the transcription initiation site [5]; in this context, it is important to note that the MyoD response element is located at different positions in rats compared to mice/humans, which may cause species-specific differences in UCP3 expression [14]. The expressing level of uncoupling protein 3 (ucp3) gene in Guanling cattle different tissues. Mpared to that for pGL3-promoter-ucp3-pro/pGL3-basic (Figure 5). These results indicate that the additional promoter had a positive affect on the expression of the target gene before UCP3 promoter. MRFs family and MEF2A potential activate an UCP3 promoter fragment of 1080 bp were detecMtedRFtshrfaomugilhy danudalMluEcFif2eArapseotreenptioarltaerctgiveantee aasnsUayCaPf3teprrocomtroatenrsfferactgimone.nTt oofe1v0a8l0uabtpe wtheereindfeluteecnteced othf rtohuegMh RdFusalfalumciilfyeraansde rMepEoFr2tAer wgeenceomaspsaayreadftceortrcaontrsafencstfieocntsiown.ithToorevwaliuthaoteuttheexpinreflsusieonnceveocftothrse fMorRMFsyfoaDm,ilMy yafn5d, MMyEfF62, AMwyoeGcoamndpaMredEFc2oAtrafnascfteocrtsioknnsowwinthtoor rwegituhloautet eUxCprPe3sspiornomveoctteorrsexfoprreMssyiooDn, (MFiygfu5r,eM8)y.f6In, MthyeopGGaLn3d-baMsiEc-Fu2cAp3f-apc1totrrasnksfneoctwedn gtroourepg,uthlaetepGULC3P-b3aspicrovmecototerrshexopwreedssrieosnpo(Fnisgivuernees8s). tIontthheecpoG-eLxp3r-besassiiocn-uocfpM3-ypf15,trManyfs6feocrteMdygorDouaplo, ntheerepsGpLec3t-ibvaesliyc. vFeocrtoprGsLh3o-wbaesdicr-uescppo3n-ps1iv, ethneesascttoivtihtye icnoc-reexapsreedss1io.4n-foofldMmyfo5,reMbyyf6MoryMf5,y3o.D3-afololdnemreosrpeebcytivMelyyf.6F, oarnpdG2L-f3o-lbdasmico-ruecpb3y-pM1y, othDe, accotmivpitayriendcrteoatsheed a1c.4ti-vfoitlidesminorCe2bCy1M2 cyef5ll,s3t.r3a-nfoslfdecmteodrewbityhMpGyfL6,3a-Bnadsi2c-.fTohldisminodriecabtyesMthyaotDt,hceoimncpraeraesdestoinththeeacatcitvivitiiteys oinf tChe2CU1C2Pc3epllrsomtraontesrfeacrteebdrowuigthhtpaGboLu3t-Bbaystihce.TthraisnsicnrdipictaiotensfathctaotrsthMeyifn5c,rMeaysf6esanind tMheyoaDc.tiIvnitcyonotfratshte, tUhCe Pc3o-pexropmreossteiornaroef bMroyuogGhat nadboMutEbFy2Athreestrualntesdcriinptioonnlyfa0c.t3o7r-saMndyf50,.3M2-yfof6ldancdhaMngyeosDo. fInluccoifnetrraasset, athcteivcioty-e, xrepsrpeesscitoivneloyf.TMheysoeGreasnudltMs inEdF2icAateretshualtteMdyionGoannldy M0.3E7F-2aAnhdav0.e3a2-nfoelgdatcihveanrgegeus loaftioluncrifoelreaosef UacCtiPv3itpy,rroemspoetecrti.vely.These results indicate that MyoG and MEF2A have a negative regulation role of UCP3 promoter
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