Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion
Abstract Background Progression of prostate cancer from benign local tumors to metastatic carcinomas is a multistep process. Here we have investigated the signaling pathways that support migration and invasion of prostate cancer cells, focusing on the role of the NFATC1 transcription factor and its...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-11-01
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| Series: | Cell Communication and Signaling |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12964-019-0463-y |
| _version_ | 1819021314998927360 |
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| author | Sini K. Eerola Niina M. Santio Sanni Rinne Petri Kouvonen Garry L. Corthals Mauro Scaravilli Giovanni Scala Angela Serra Dario Greco Pekka Ruusuvuori Leena Latonen Eeva-Marja Rainio Tapio Visakorpi Päivi J. Koskinen |
| author_facet | Sini K. Eerola Niina M. Santio Sanni Rinne Petri Kouvonen Garry L. Corthals Mauro Scaravilli Giovanni Scala Angela Serra Dario Greco Pekka Ruusuvuori Leena Latonen Eeva-Marja Rainio Tapio Visakorpi Päivi J. Koskinen |
| author_sort | Sini K. Eerola |
| collection | DOAJ |
| description | Abstract Background Progression of prostate cancer from benign local tumors to metastatic carcinomas is a multistep process. Here we have investigated the signaling pathways that support migration and invasion of prostate cancer cells, focusing on the role of the NFATC1 transcription factor and its post-translational modifications. We have previously identified NFATC1 as a substrate for the PIM1 kinase and shown that PIM1-dependent phosphorylation increases NFATC1 activity without affecting its subcellular localization. Both PIM kinases and NFATC1 have been reported to promote cancer cell migration, invasion and angiogenesis, but it has remained unclear whether the effects of NFATC1 are phosphorylation-dependent and which downstream targets are involved. Methods We used mass spectrometry to identify PIM1 phosphorylation target sites in NFATC1, and analysed their functional roles in three prostate cancer cell lines by comparing phosphodeficient mutants to wild-type NFATC1. We used luciferase assays to determine effects of phosphorylation on NFAT-dependent transcriptional activity, and migration and invasion assays to evaluate effects on cell motility. We also performed a microarray analysis to identify novel PIM1/NFATC1 targets, and validated one of them with both cellular expression analyses and in silico in clinical prostate cancer data sets. Results Here we have identified ten PIM1 target sites in NFATC1 and found that prevention of their phosphorylation significantly decreases the transcriptional activity as well as the pro-migratory and pro-invasive effects of NFATC1 in prostate cancer cells. We observed that also PIM2 and PIM3 can phosphorylate NFATC1, and identified several novel putative PIM1/NFATC1 target genes. These include the ITGA5 integrin, which is differentially expressed in the presence of wild-type versus phosphorylation-deficient NFATC1, and which is coexpressed with PIM1 and NFATC1 in clinical prostate cancer specimens. Conclusions Based on our data, phosphorylation of PIM1 target sites stimulates NFATC1 activity and enhances its ability to promote prostate cancer cell migration and invasion. Therefore, inhibition of the interplay between PIM kinases and NFATC1 may have therapeutic implications for patients with metastatic forms of cancer. Graphical abstract |
| first_indexed | 2024-12-21T04:05:09Z |
| format | Article |
| id | doaj.art-a0efde44d00b4da78c2fb9399ca6eec3 |
| institution | Directory Open Access Journal |
| issn | 1478-811X |
| language | English |
| last_indexed | 2024-12-21T04:05:09Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell Communication and Signaling |
| spelling | doaj.art-a0efde44d00b4da78c2fb9399ca6eec32022-12-21T19:16:37ZengBMCCell Communication and Signaling1478-811X2019-11-0117111610.1186/s12964-019-0463-yPhosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasionSini K. Eerola0Niina M. Santio1Sanni Rinne2Petri Kouvonen3Garry L. Corthals4Mauro Scaravilli5Giovanni Scala6Angela Serra7Dario Greco8Pekka Ruusuvuori9Leena Latonen10Eeva-Marja Rainio11Tapio Visakorpi12Päivi J. Koskinen13Department of Biology, University of TurkuDepartment of Biology, University of TurkuDepartment of Biology, University of TurkuTurku Centre for Biotechnology, University of TurkuTurku Centre for Biotechnology, University of TurkuFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalDepartment of Biology, University of TurkuFaculty of Medicine and Health Technology, Tampere University and Tays Cancer Center, Tampere University HospitalDepartment of Biology, University of TurkuAbstract Background Progression of prostate cancer from benign local tumors to metastatic carcinomas is a multistep process. Here we have investigated the signaling pathways that support migration and invasion of prostate cancer cells, focusing on the role of the NFATC1 transcription factor and its post-translational modifications. We have previously identified NFATC1 as a substrate for the PIM1 kinase and shown that PIM1-dependent phosphorylation increases NFATC1 activity without affecting its subcellular localization. Both PIM kinases and NFATC1 have been reported to promote cancer cell migration, invasion and angiogenesis, but it has remained unclear whether the effects of NFATC1 are phosphorylation-dependent and which downstream targets are involved. Methods We used mass spectrometry to identify PIM1 phosphorylation target sites in NFATC1, and analysed their functional roles in three prostate cancer cell lines by comparing phosphodeficient mutants to wild-type NFATC1. We used luciferase assays to determine effects of phosphorylation on NFAT-dependent transcriptional activity, and migration and invasion assays to evaluate effects on cell motility. We also performed a microarray analysis to identify novel PIM1/NFATC1 targets, and validated one of them with both cellular expression analyses and in silico in clinical prostate cancer data sets. Results Here we have identified ten PIM1 target sites in NFATC1 and found that prevention of their phosphorylation significantly decreases the transcriptional activity as well as the pro-migratory and pro-invasive effects of NFATC1 in prostate cancer cells. We observed that also PIM2 and PIM3 can phosphorylate NFATC1, and identified several novel putative PIM1/NFATC1 target genes. These include the ITGA5 integrin, which is differentially expressed in the presence of wild-type versus phosphorylation-deficient NFATC1, and which is coexpressed with PIM1 and NFATC1 in clinical prostate cancer specimens. Conclusions Based on our data, phosphorylation of PIM1 target sites stimulates NFATC1 activity and enhances its ability to promote prostate cancer cell migration and invasion. Therefore, inhibition of the interplay between PIM kinases and NFATC1 may have therapeutic implications for patients with metastatic forms of cancer. Graphical abstracthttp://link.springer.com/article/10.1186/s12964-019-0463-yProstate cancerMetastatic carcinomaNFATC1PIM kinasesCell motility |
| spellingShingle | Sini K. Eerola Niina M. Santio Sanni Rinne Petri Kouvonen Garry L. Corthals Mauro Scaravilli Giovanni Scala Angela Serra Dario Greco Pekka Ruusuvuori Leena Latonen Eeva-Marja Rainio Tapio Visakorpi Päivi J. Koskinen Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion Cell Communication and Signaling Prostate cancer Metastatic carcinoma NFATC1 PIM kinases Cell motility |
| title | Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| title_full | Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| title_fullStr | Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| title_full_unstemmed | Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| title_short | Phosphorylation of NFATC1 at PIM1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| title_sort | phosphorylation of nfatc1 at pim1 target sites is essential for its ability to promote prostate cancer cell migration and invasion |
| topic | Prostate cancer Metastatic carcinoma NFATC1 PIM kinases Cell motility |
| url | http://link.springer.com/article/10.1186/s12964-019-0463-y |
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