XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress
Abstract Background Centromere protein F (CENPF) is a key component of the kinetochore complex involved in mitosis, cell differentiation and cellular response to stresses. However, the alteration of CENPF in response to endoplasmic reticulum (ER) stress has not been well described. In the present st...
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| Format: | Article |
| Language: | English |
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BMC
2020-09-01
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| Series: | Cancer Cell International |
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| Online Access: | http://link.springer.com/article/10.1186/s12935-020-01553-9 |
| _version_ | 1818113785574981632 |
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| author | Tao Shen Yan Li Shuang Liang Zhiguang Chen |
| author_facet | Tao Shen Yan Li Shuang Liang Zhiguang Chen |
| author_sort | Tao Shen |
| collection | DOAJ |
| description | Abstract Background Centromere protein F (CENPF) is a key component of the kinetochore complex involved in mitosis, cell differentiation and cellular response to stresses. However, the alteration of CENPF in response to endoplasmic reticulum (ER) stress has not been well described. In the present study, we investigate CENPF regulation in response to ER stress. Methods Quantitative real-time polymerase chain reaction and western blotting were used to determine CENPF expression under ER stress. Luciferase activity analysis was performed to investigate the promoter regions contributing to CENPF transcription in response to TG. Chromatin immunoprecipitation (ChIP) and ChIP Re-IP assays were used to determine if X-box binding protein 1 (XBP1) and/or activating transcription factor 6α (ATF6α) bind in the CENPF promoter region. Cell apoptosis and proliferation were analyzed using TUNEL, cell growth and clonogenic assays. Results CENPF expression is dramatically reduced under ER stress induced by thapsigargin (TG), brefeldin A (BFA), or tunicamycin (TM) and this downregulation of CENPF expression was dependent on XBP1 and ATF6α. Luciferase activity analysis of the truncated CENPF promoter indicates that regions from bases − 679 to − 488 and from − 241 to − 78 in the CENPF promoter were sensitive to TG treatment. Additionally, ChIP and ChIP Re-IP assays reveal that XBP1 and ATF6α were assembled on the same regions of CENPF promoter. Notably, we identify two XBP1 binding sequences at positions − 567 and − 192, to which XBP1 binding was enhanced by TG. Finally, CENPF overexpression inhibits cell apoptosis and promotes cell proliferation in response to ER stress. Conclusion In summary, these results demonstrate that ER stress plays a crucial role in CENPF expression, and XBP1 may up-regulate DNA-binding affinities after TG treatment to the promoter of CENPF. These findings may contribute to the understanding of the molecular mechanism of CENPF regulation. |
| first_indexed | 2024-12-11T03:40:21Z |
| format | Article |
| id | doaj.art-4093ec1e94504e4d9087c6ff8fe23b44 |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-12-11T03:40:21Z |
| publishDate | 2020-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-4093ec1e94504e4d9087c6ff8fe23b442022-12-22T01:22:09ZengBMCCancer Cell International1475-28672020-09-0120111110.1186/s12935-020-01553-9XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stressTao Shen0Yan Li1Shuang Liang2Zhiguang Chen3Department of Orthopedics, Shengjing Hospital of China Medical UniversityDepartment of Cell Biology, Key Laboratory of Cell Biology, Ministry of Public Health and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical UniversityDepartment of Laboratory Medicine and Pathology, University of MinnesotaDepartment of Orthopedics, Shengjing Hospital of China Medical UniversityAbstract Background Centromere protein F (CENPF) is a key component of the kinetochore complex involved in mitosis, cell differentiation and cellular response to stresses. However, the alteration of CENPF in response to endoplasmic reticulum (ER) stress has not been well described. In the present study, we investigate CENPF regulation in response to ER stress. Methods Quantitative real-time polymerase chain reaction and western blotting were used to determine CENPF expression under ER stress. Luciferase activity analysis was performed to investigate the promoter regions contributing to CENPF transcription in response to TG. Chromatin immunoprecipitation (ChIP) and ChIP Re-IP assays were used to determine if X-box binding protein 1 (XBP1) and/or activating transcription factor 6α (ATF6α) bind in the CENPF promoter region. Cell apoptosis and proliferation were analyzed using TUNEL, cell growth and clonogenic assays. Results CENPF expression is dramatically reduced under ER stress induced by thapsigargin (TG), brefeldin A (BFA), or tunicamycin (TM) and this downregulation of CENPF expression was dependent on XBP1 and ATF6α. Luciferase activity analysis of the truncated CENPF promoter indicates that regions from bases − 679 to − 488 and from − 241 to − 78 in the CENPF promoter were sensitive to TG treatment. Additionally, ChIP and ChIP Re-IP assays reveal that XBP1 and ATF6α were assembled on the same regions of CENPF promoter. Notably, we identify two XBP1 binding sequences at positions − 567 and − 192, to which XBP1 binding was enhanced by TG. Finally, CENPF overexpression inhibits cell apoptosis and promotes cell proliferation in response to ER stress. Conclusion In summary, these results demonstrate that ER stress plays a crucial role in CENPF expression, and XBP1 may up-regulate DNA-binding affinities after TG treatment to the promoter of CENPF. These findings may contribute to the understanding of the molecular mechanism of CENPF regulation.http://link.springer.com/article/10.1186/s12935-020-01553-9ER stressCENPFExpression regulationXBP1ATF6α |
| spellingShingle | Tao Shen Yan Li Shuang Liang Zhiguang Chen XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress Cancer Cell International ER stress CENPF Expression regulation XBP1 ATF6α |
| title | XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress |
| title_full | XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress |
| title_fullStr | XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress |
| title_full_unstemmed | XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress |
| title_short | XBP1 negatively regulates CENPF expression via recruiting ATF6α to the promoter during ER stress |
| title_sort | xbp1 negatively regulates cenpf expression via recruiting atf6α to the promoter during er stress |
| topic | ER stress CENPF Expression regulation XBP1 ATF6α |
| url | http://link.springer.com/article/10.1186/s12935-020-01553-9 |
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