PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation
Abstract Yes-associated protein (YAP) is a transcriptional coactivator that is essential for the malignancy of various cancers. We have previously shown that YAP activity is positively regulated by phosphatidylserine (PS) in recycling endosomes (REs). However, the mechanism by which YAP is activated...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-47138-0 |
| _version_ | 1797559914662461440 |
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| author | Chiaki Inoue Kojiro Mukai Tatsuyuki Matsudaira Jun Nakayama Nozomu Kono Junken Aoki Hiroyuki Arai Yasunori Uchida Tomohiko Taguchi |
| author_facet | Chiaki Inoue Kojiro Mukai Tatsuyuki Matsudaira Jun Nakayama Nozomu Kono Junken Aoki Hiroyuki Arai Yasunori Uchida Tomohiko Taguchi |
| author_sort | Chiaki Inoue |
| collection | DOAJ |
| description | Abstract Yes-associated protein (YAP) is a transcriptional coactivator that is essential for the malignancy of various cancers. We have previously shown that YAP activity is positively regulated by phosphatidylserine (PS) in recycling endosomes (REs). However, the mechanism by which YAP is activated by PS in REs remains unknown. In the present study, we examined a group of protein phosphatases (11 phosphatases) that we had identified previously as PS-proximity protein candidates. Knockdown experiments of these phosphatases suggested that PPP1R12A, a regulatory subunit of the myosin phosphatase complex, was essential for YAP-dependent proliferation of triple-negative breast cancer MDA-MB-231 cells. Knockdown of PPP1R12A increased the level of phosphorylated YAP, reduced that of YAP in the nucleus, and suppressed the transcription of CTGF (a YAP-regulated gene), reinforcing the role of PPP1R12A in YAP activation. ATP8A1 is a PS-flippase that concentrates PS in the cytosolic leaflet of the RE membrane and positively regulates YAP signalling. In subcellular fractionation experiments using cell lysates, PPP1R12A in control cells was recovered exclusively in the microsomal fraction. In contrast, a fraction of PPP1R12A in ATP8A1-depleted cells was recovered in the cytosolic fraction. Cohort data available from the Cancer Genome Atlas showed that high expression of PPP1R12A, PP1B encoding the catalytic subunit of the myosin phosphatase complex, or ATP8A1 correlated with poor prognosis in breast cancer patients. These results suggest that the “ATP8A1-PS-YAP phosphatase” axis in REs facilitates YAP activation and thus cell proliferation. |
| first_indexed | 2024-03-10T17:51:56Z |
| format | Article |
| id | doaj.art-8d6cacaebed848179498734751cbfd5d |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-10T17:51:56Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-8d6cacaebed848179498734751cbfd5d2023-11-20T09:19:11ZengNature PortfolioScientific Reports2045-23222023-11-0113111010.1038/s41598-023-47138-0PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activationChiaki Inoue0Kojiro Mukai1Tatsuyuki Matsudaira2Jun Nakayama3Nozomu Kono4Junken Aoki5Hiroyuki Arai6Yasunori Uchida7Tomohiko Taguchi8Department of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of TokyoLaboratory of Organelle Pathophysiology, Department of Integrative Life Sciences, Graduate School of Life Sciences, Tohoku UniversityDepartment of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of TokyoLaboratory of Integrative Oncology, National Cancer Center Research InstituteDepartment of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of TokyoDepartment of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of TokyoDepartment of Health Chemistry, Graduate School of Pharmaceutical Sciences, University of TokyoLaboratory of Organelle Pathophysiology, Department of Integrative Life Sciences, Graduate School of Life Sciences, Tohoku UniversityLaboratory of Organelle Pathophysiology, Department of Integrative Life Sciences, Graduate School of Life Sciences, Tohoku UniversityAbstract Yes-associated protein (YAP) is a transcriptional coactivator that is essential for the malignancy of various cancers. We have previously shown that YAP activity is positively regulated by phosphatidylserine (PS) in recycling endosomes (REs). However, the mechanism by which YAP is activated by PS in REs remains unknown. In the present study, we examined a group of protein phosphatases (11 phosphatases) that we had identified previously as PS-proximity protein candidates. Knockdown experiments of these phosphatases suggested that PPP1R12A, a regulatory subunit of the myosin phosphatase complex, was essential for YAP-dependent proliferation of triple-negative breast cancer MDA-MB-231 cells. Knockdown of PPP1R12A increased the level of phosphorylated YAP, reduced that of YAP in the nucleus, and suppressed the transcription of CTGF (a YAP-regulated gene), reinforcing the role of PPP1R12A in YAP activation. ATP8A1 is a PS-flippase that concentrates PS in the cytosolic leaflet of the RE membrane and positively regulates YAP signalling. In subcellular fractionation experiments using cell lysates, PPP1R12A in control cells was recovered exclusively in the microsomal fraction. In contrast, a fraction of PPP1R12A in ATP8A1-depleted cells was recovered in the cytosolic fraction. Cohort data available from the Cancer Genome Atlas showed that high expression of PPP1R12A, PP1B encoding the catalytic subunit of the myosin phosphatase complex, or ATP8A1 correlated with poor prognosis in breast cancer patients. These results suggest that the “ATP8A1-PS-YAP phosphatase” axis in REs facilitates YAP activation and thus cell proliferation.https://doi.org/10.1038/s41598-023-47138-0 |
| spellingShingle | Chiaki Inoue Kojiro Mukai Tatsuyuki Matsudaira Jun Nakayama Nozomu Kono Junken Aoki Hiroyuki Arai Yasunori Uchida Tomohiko Taguchi PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation Scientific Reports |
| title | PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation |
| title_full | PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation |
| title_fullStr | PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation |
| title_full_unstemmed | PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation |
| title_short | PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation |
| title_sort | ppp1r12a is a recycling endosomal phosphatase that facilitates yap activation |
| url | https://doi.org/10.1038/s41598-023-47138-0 |
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