Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP
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 reti...
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| Format: | Article |
| Language: | English |
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China Science Publishing & Media Ltd.
2022-05-01
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| Series: | Acta Biochimica et Biophysica Sinica |
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| Online Access: | https://www.sciengine.com/doi/10.3724/abbs.2022041 |
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| author | Fan Guangchun Tao Zhengxin Chen Shang Zhang Huahua Yu Rongjie |
| author_facet | Fan Guangchun Tao Zhengxin Chen Shang Zhang Huahua Yu Rongjie |
| author_sort | Fan Guangchun |
| collection | DOAJ |
| description | 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 MPP-induced cell model of Parkinson<?A3B2 tf="TT5843c571"?>’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. |
| format | Article |
| id | doaj-art-0293c3fd22f14434a5f661ffabf67374 |
| institution | OA Journals |
| issn | 1672-9145 |
| language | English |
| publishDate | 2022-05-01 |
| publisher | China Science Publishing & Media Ltd. |
| record_format | Article |
| series | Acta Biochimica et Biophysica Sinica |
| spelling | doaj-art-0293c3fd22f14434a5f661ffabf673742025-08-20T02:26:28ZengChina Science Publishing & Media Ltd.Acta Biochimica et Biophysica Sinica1672-91452022-05-015465767210.3724/abbs.202204120d259ccPositive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAPFan Guangchun0Tao Zhengxin1Chen Shang2Zhang Huahua3Yu Rongjie4["Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou 510630, China"]["Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou 510630, China"]["Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou 510630, China"]["Department of Medical Genetics, Guangdong Medical University, Dongguan 523000, China"]["Department of Cell Biology, College of Life Science and Technology, Jinan University, Guangzhou 510630, China","Guangdong Province Key Laboratory of Bioengineering Medicine, Guangzhou 510630, China","Guangdong Provincial Biotechnology Drug & Engineering Technology Research Center, Guangzhou 510630, China","National Engineering Research Center of Genetic Medicine, Guangzhou 510630, China"]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 MPP-induced cell model of Parkinson<?A3B2 tf="TT5843c571"?>’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.https://www.sciengine.com/doi/10.3724/abbs.2022041PACAPPAC1-Rpositive allosteric modulationnuclear translocation |
| spellingShingle | Fan Guangchun Tao Zhengxin Chen Shang Zhang Huahua Yu Rongjie Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP Acta Biochimica et Biophysica Sinica PACAP PAC1-R positive allosteric modulation nuclear translocation |
| title | Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP |
| title_full | Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP |
| title_fullStr | Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP |
| title_full_unstemmed | Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP |
| title_short | Positive allosteric regulation of PAC1-R up-regulates<?A3B2 ACK?>PAC1-R and its specific ligand PACAP |
| title_sort | positive allosteric regulation of pac1 r up regulates a3b2 ack pac1 r and its specific ligand pacap |
| topic | PACAP PAC1-R positive allosteric modulation nuclear translocation |
| url | https://www.sciengine.com/doi/10.3724/abbs.2022041 |
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