Positive allosteric regulation of PAC1-R up-regulates PAC1-R and its specific ligand PACAP  

作　　者：Guangchun Fan Zhengxin Tao Shang Chen Huahua Zhang Rongjie Yu 

机构地区：[1]Department of Cell Biology,College of Life Science and Technology,Jinan University,Guangzhou 510630,China [2]Guangdong Province Key Laboratory of Bioengineering Medicine,Guangzhou 510630,China [3]Guangdong Provincial Biotechnology Drug&Engineering Technology Research Center,Guangzhou 510630,China [4]National Engineering Research Center of Genetic Medicine,Guangzhou 510630,China [5]Department of Medical Genetics,Guangdong Medical University,Dongguan 523000,China

出　　处：《Acta Biochimica et Biophysica Sinica》2022年第5期657-672,共16页生物化学与生物物理学报（英文版）

基　　金：supported by the grants from the National Natural Science Foundation of China(Nos.31100545 and 31670848);the Natural Science Foundation of Guangdong Province(Nos.2016A030313087 and 2022A1515011158).

摘　　要：PAC1-R is a recognized preferential receptor for the neuropeptide of pituitary adenylate cyclase-activating polypeptide(PACAP),which mediates neuroprotective and nerve regenerative activities of PACAP.In this study,we found that in both PAC1R-CHO cells with high expression of PAC1R-eGFP and retinal ganglion cells(RGC-5)with the natural expression of PAC1-R,oligo-peptide PACAP(28-38)and the positively charged arginine-rich penetrating peptide TAT,as positive allosteric modulators of PAC1-R,significantly trigger the nuclear translocation of PAC1-R.The chromatin immunoprecipitation(ChIP)-PCR results show that the nuclear translocated PAC1-R binds with the promoter regions of PAC1-R and its specific ligand PACAP.The up-regulated promoter activities of PAC1-R and PACAP induced by PACAP(28-38)or TAT are positively correlative with the increase of the expression levels of PAC1-R and PACAP.Moreover,the nuclear translocation of PAC1-R induced by PACAP(28-38)or TAT is significantly inhibited by the mutation of PAC1-R on Cys25 and the palmitoylation inhibitor 2-bromopalmitate.Meanwhile,the increase in both PAC1-R and PACAP levels and the neuroprotective activities of PACAP(28-38)and TAT in MPPinduced cell model of Parkinson’s disease are synchronously inhibited by 2-bromopalmitate,which are positively correlated with the nuclear translocation of PAC1-R induced by PACAP(28-38)or TAT.Bioinformatics analysis and motif enrichment analysis following ChIP-sequencing show that the transcription factors including SP1,Zic2,GATA1,REST and YY1 may be recruited by nuclear PAC1-R and involved in regulating the promoter activities of PAC1-R and PACAP.ChIP-sequencing and related bioinformatics analysis show that the downstream target genes regulated by the nuclear PAC1-R are mostly involved in the process of cellular stress and related to neuroprotection,neuronal genesis and development.

关 键 词：PACAP PAC1-R positive allosteric modulation nuclear translocation 

分 类 号：O62[理学—有机化学]