KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA
Abstract Aims To explore the function of phosphorylation of KAP1 (p-KAP1) at the serine-824 site (S824) in the proliferation and apoptosis of endogenous neural stem cells (NSCs) after cerebral ischemic/reperfusion (I/R). Methods The apoptosis and proliferation of C17.2 cells transfected with the p-K...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-07-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02962-5 |
| _version_ | 1811219284972535808 |
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| author | Wan Wang Tianqing Yan Xinjian Guo Heng Cai Chang Liang Linyan Huang Yanling Wang Ping Ma Suhua Qi |
| author_facet | Wan Wang Tianqing Yan Xinjian Guo Heng Cai Chang Liang Linyan Huang Yanling Wang Ping Ma Suhua Qi |
| author_sort | Wan Wang |
| collection | DOAJ |
| description | Abstract Aims To explore the function of phosphorylation of KAP1 (p-KAP1) at the serine-824 site (S824) in the proliferation and apoptosis of endogenous neural stem cells (NSCs) after cerebral ischemic/reperfusion (I/R). Methods The apoptosis and proliferation of C17.2 cells transfected with the p-KAP1-expression plasmids and the expression of proliferation cell nuclear antigen (PCNA) and p-KAP1 were detected by immunofluorescence and Western blotting after the Oxygen Glucose deprivation/reperfusion model (OGD/R). The interaction of p-KAP1 and CUL4A with PCNA was analyzed by immunoprecipitation. In the rats MCAO model, we performed the adeno-associated virus (AAV) 2/9 gene delivery of p-KAP1 mutants to verify the proliferation of endogenous NSCs and the colocalization of PCNA and CUL4A by immunofluorescence. Results The level of p-KAP1 was significantly down-regulated in the stroke model in vivo and in vitro. Simulated p-KAP1(S824) significantly increased the proliferation of C17.2 cells and the expression of PCNA after OGD/R. Simulated p-KAP1(S824) enhanced the binding of p-KAP1 and PCNA and decreased the interaction between PCNA and CUL4A in C17.2 cells subjected to OGD/R. The AAV2/9-mediated p-KAP1(S824) increased endogenous NSCs proliferation, PCNA expression, p-KAP1 binding to PCNA, and improved neurological function in the rat MCAO model. Conclusions Our findings confirmed that simulated p-KAP1(S824) improved the survival and proliferation of endogenous NSCs. The underlying mechanism is that highly expressed p-KAP1(S824) promotes binding to PCNA, and inhibits the binding of CUL4A to PCNA. This reduced CUL4A-mediated ubiquitination degradation to increase the stability of PCNA and promote the survival and proliferation of NSCs. |
| first_indexed | 2024-04-12T07:23:34Z |
| format | Article |
| id | doaj.art-4d7efd530c2046c18246747c238f6701 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-12T07:23:34Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-4d7efd530c2046c18246747c238f67012022-12-22T03:42:15ZengBMCStem Cell Research & Therapy1757-65122022-07-0113111810.1186/s13287-022-02962-5KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNAWan Wang0Tianqing Yan1Xinjian Guo2Heng Cai3Chang Liang4Linyan Huang5Yanling Wang6Ping Ma7Suhua Qi8School of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Basic Medical Science, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversitySchool of Medical Technology, Xuzhou Key Laboratory of Laboratory Diagnostics, Xuzhou Medical UniversityAbstract Aims To explore the function of phosphorylation of KAP1 (p-KAP1) at the serine-824 site (S824) in the proliferation and apoptosis of endogenous neural stem cells (NSCs) after cerebral ischemic/reperfusion (I/R). Methods The apoptosis and proliferation of C17.2 cells transfected with the p-KAP1-expression plasmids and the expression of proliferation cell nuclear antigen (PCNA) and p-KAP1 were detected by immunofluorescence and Western blotting after the Oxygen Glucose deprivation/reperfusion model (OGD/R). The interaction of p-KAP1 and CUL4A with PCNA was analyzed by immunoprecipitation. In the rats MCAO model, we performed the adeno-associated virus (AAV) 2/9 gene delivery of p-KAP1 mutants to verify the proliferation of endogenous NSCs and the colocalization of PCNA and CUL4A by immunofluorescence. Results The level of p-KAP1 was significantly down-regulated in the stroke model in vivo and in vitro. Simulated p-KAP1(S824) significantly increased the proliferation of C17.2 cells and the expression of PCNA after OGD/R. Simulated p-KAP1(S824) enhanced the binding of p-KAP1 and PCNA and decreased the interaction between PCNA and CUL4A in C17.2 cells subjected to OGD/R. The AAV2/9-mediated p-KAP1(S824) increased endogenous NSCs proliferation, PCNA expression, p-KAP1 binding to PCNA, and improved neurological function in the rat MCAO model. Conclusions Our findings confirmed that simulated p-KAP1(S824) improved the survival and proliferation of endogenous NSCs. The underlying mechanism is that highly expressed p-KAP1(S824) promotes binding to PCNA, and inhibits the binding of CUL4A to PCNA. This reduced CUL4A-mediated ubiquitination degradation to increase the stability of PCNA and promote the survival and proliferation of NSCs.https://doi.org/10.1186/s13287-022-02962-5Cerebral ischemia/reperfusion (I/R)Neural stem cells (NSCs)ProliferationKRAB domain protein 1(KAP1)PCNA |
| spellingShingle | Wan Wang Tianqing Yan Xinjian Guo Heng Cai Chang Liang Linyan Huang Yanling Wang Ping Ma Suhua Qi KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA Stem Cell Research & Therapy Cerebral ischemia/reperfusion (I/R) Neural stem cells (NSCs) Proliferation KRAB domain protein 1(KAP1) PCNA |
| title | KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA |
| title_full | KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA |
| title_fullStr | KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA |
| title_full_unstemmed | KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA |
| title_short | KAP1 phosphorylation promotes the survival of neural stem cells after ischemia/reperfusion by maintaining the stability of PCNA |
| title_sort | kap1 phosphorylation promotes the survival of neural stem cells after ischemia reperfusion by maintaining the stability of pcna |
| topic | Cerebral ischemia/reperfusion (I/R) Neural stem cells (NSCs) Proliferation KRAB domain protein 1(KAP1) PCNA |
| url | https://doi.org/10.1186/s13287-022-02962-5 |
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