Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity
Willin/FRMD6 is part of a family of proteins with a 4.1 ezrin-radixin-moesin (FERM) domain. It has been identified as an upstream activator of the Hippo pathway and, when aberrant in its expression, is associated with human diseases and disorders. Even though Willin/FRMD6 was originally discovered i...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-09-01
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| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fncel.2020.552213/full |
| _version_ | 1818996892598534144 |
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| author | Nils M. Kronenberg Nils M. Kronenberg Andrew Tilston-Lunel Andrew Tilston-Lunel Frances E. Thompson Doris Chen Wanjia Yu Kishan Dholakia Kishan Dholakia Malte C. Gather Malte C. Gather Frank J. Gunn-Moore |
| author_facet | Nils M. Kronenberg Nils M. Kronenberg Andrew Tilston-Lunel Andrew Tilston-Lunel Frances E. Thompson Doris Chen Wanjia Yu Kishan Dholakia Kishan Dholakia Malte C. Gather Malte C. Gather Frank J. Gunn-Moore |
| author_sort | Nils M. Kronenberg |
| collection | DOAJ |
| description | Willin/FRMD6 is part of a family of proteins with a 4.1 ezrin-radixin-moesin (FERM) domain. It has been identified as an upstream activator of the Hippo pathway and, when aberrant in its expression, is associated with human diseases and disorders. Even though Willin/FRMD6 was originally discovered in the rat sciatic nerve, most studies have focused on its functional roles in cells outside of the nervous system, where Willin/FRMD6 is involved in the formation of apical junctional cell-cell complexes and in regulating cell migration. Here, we investigate the biochemical and biophysical role of Willin/FRMD6 in neuronal cells, employing the commonly used SH-SY5Y neuronal model cell system and combining biochemical measurements with Elastic Resonator Interference Stress Micropscopy (ERISM). We present the first direct evidence that Willin/FRMD6 expression influences both the cell mechanical phenotype and neuronal differentiation. By investigating cells with increased and decreased Willin/FRMD6 expression levels, we show that Willin/FRMD6 not only affects proliferation and migration capacity of cells but also leads to changes in cell morphology and an enhanced formation of neurite-like membrane extensions. These changes were accompanied by alterations of biophysical parameters such as cell force, the organization of actin stress fibers and the formation of focal adhesions. At the biochemical level, changes in Willin/FRMD6 expression inversely affected the activity of the extracellular signal-regulated kinases (ERK) pathway and downstream transcriptional factor NeuroD1, which seems to prime SH-SY5Y cells for retinoic acid (RA)-induced neuronal differentiation. |
| first_indexed | 2024-12-20T21:36:58Z |
| format | Article |
| id | doaj.art-810fe848f8e84347b5596fd3f2b2ab06 |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-12-20T21:36:58Z |
| publishDate | 2020-09-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-810fe848f8e84347b5596fd3f2b2ab062022-12-21T19:25:55ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022020-09-011410.3389/fncel.2020.552213552213Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 ActivityNils M. Kronenberg0Nils M. Kronenberg1Andrew Tilston-Lunel2Andrew Tilston-Lunel3Frances E. Thompson4Doris Chen5Wanjia Yu6Kishan Dholakia7Kishan Dholakia8Malte C. Gather9Malte C. Gather10Frank J. Gunn-Moore11Centre of Biophotonics and SUPA, School of Physics and Astronomy, University of St Andrews, St Andrews, United KingdomCentre for Nanobiophotonics, Department of Chemistry, University of Cologne, Cologne, GermanyCentre of Biophotonics, School of Biology, University of St Andrews, St Andrews, United KingdomDepartment of Biochemistry, School of Medicine, Boston University, Boston, MA, United StatesCentre of Biophotonics and SUPA, School of Physics and Astronomy, University of St Andrews, St Andrews, United KingdomCentre of Biophotonics, School of Biology, University of St Andrews, St Andrews, United KingdomCentre of Biophotonics, School of Biology, University of St Andrews, St Andrews, United KingdomCentre of Biophotonics and SUPA, School of Physics and Astronomy, University of St Andrews, St Andrews, United KingdomDepartment of Physics, College of Science, Yonsei University, Seoul, South KoreaCentre of Biophotonics and SUPA, School of Physics and Astronomy, University of St Andrews, St Andrews, United KingdomCentre for Nanobiophotonics, Department of Chemistry, University of Cologne, Cologne, GermanyCentre of Biophotonics, School of Biology, University of St Andrews, St Andrews, United KingdomWillin/FRMD6 is part of a family of proteins with a 4.1 ezrin-radixin-moesin (FERM) domain. It has been identified as an upstream activator of the Hippo pathway and, when aberrant in its expression, is associated with human diseases and disorders. Even though Willin/FRMD6 was originally discovered in the rat sciatic nerve, most studies have focused on its functional roles in cells outside of the nervous system, where Willin/FRMD6 is involved in the formation of apical junctional cell-cell complexes and in regulating cell migration. Here, we investigate the biochemical and biophysical role of Willin/FRMD6 in neuronal cells, employing the commonly used SH-SY5Y neuronal model cell system and combining biochemical measurements with Elastic Resonator Interference Stress Micropscopy (ERISM). We present the first direct evidence that Willin/FRMD6 expression influences both the cell mechanical phenotype and neuronal differentiation. By investigating cells with increased and decreased Willin/FRMD6 expression levels, we show that Willin/FRMD6 not only affects proliferation and migration capacity of cells but also leads to changes in cell morphology and an enhanced formation of neurite-like membrane extensions. These changes were accompanied by alterations of biophysical parameters such as cell force, the organization of actin stress fibers and the formation of focal adhesions. At the biochemical level, changes in Willin/FRMD6 expression inversely affected the activity of the extracellular signal-regulated kinases (ERK) pathway and downstream transcriptional factor NeuroD1, which seems to prime SH-SY5Y cells for retinoic acid (RA)-induced neuronal differentiation.https://www.frontiersin.org/article/10.3389/fncel.2020.552213/fullWillin/FRMD6neuronal differentiationERK1/2cell mechanicsactinfocal adhesion |
| spellingShingle | Nils M. Kronenberg Nils M. Kronenberg Andrew Tilston-Lunel Andrew Tilston-Lunel Frances E. Thompson Doris Chen Wanjia Yu Kishan Dholakia Kishan Dholakia Malte C. Gather Malte C. Gather Frank J. Gunn-Moore Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity Frontiers in Cellular Neuroscience Willin/FRMD6 neuronal differentiation ERK1/2 cell mechanics actin focal adhesion |
| title | Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity |
| title_full | Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity |
| title_fullStr | Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity |
| title_full_unstemmed | Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity |
| title_short | Willin/FRMD6 Influences Mechanical Phenotype and Neuronal Differentiation in Mammalian Cells by Regulating ERK1/2 Activity |
| title_sort | willin frmd6 influences mechanical phenotype and neuronal differentiation in mammalian cells by regulating erk1 2 activity |
| topic | Willin/FRMD6 neuronal differentiation ERK1/2 cell mechanics actin focal adhesion |
| url | https://www.frontiersin.org/article/10.3389/fncel.2020.552213/full |
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