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PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure
The active zone of a presynaptic nerve terminal defines sites for neurotransmitter release. Its protein machinery may be organized through liquid–liquid phase separation, a mechanism for the formation of membrane-less subcellular compartments. Here, we show that the active zone protein Liprin-α3 rap...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8144191/ https://www.ncbi.nlm.nih.gov/pubmed/34031393 http://dx.doi.org/10.1038/s41467-021-23116-w |
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author | Emperador-Melero, Javier Wong, Man Yan Wang, Shan Shan H. de Nola, Giovanni Nyitrai, Hajnalka Kirchhausen, Tom Kaeser, Pascal S. |
author_facet | Emperador-Melero, Javier Wong, Man Yan Wang, Shan Shan H. de Nola, Giovanni Nyitrai, Hajnalka Kirchhausen, Tom Kaeser, Pascal S. |
author_sort | Emperador-Melero, Javier |
collection | PubMed |
description | The active zone of a presynaptic nerve terminal defines sites for neurotransmitter release. Its protein machinery may be organized through liquid–liquid phase separation, a mechanism for the formation of membrane-less subcellular compartments. Here, we show that the active zone protein Liprin-α3 rapidly and reversibly undergoes phase separation in transfected HEK293T cells. Condensate formation is triggered by Liprin-α3 PKC-phosphorylation at serine-760, and RIM and Munc13 are co-recruited into membrane-attached condensates. Phospho-specific antibodies establish phosphorylation of Liprin-α3 serine-760 in transfected cells and mouse brain tissue. In primary hippocampal neurons of newly generated Liprin-α2/α3 double knockout mice, synaptic levels of RIM and Munc13 are reduced and the pool of releasable vesicles is decreased. Re-expression of Liprin-α3 restored these presynaptic defects, while mutating the Liprin-α3 phosphorylation site to abolish phase condensation prevented this rescue. Finally, PKC activation in these neurons acutely increased RIM, Munc13 and neurotransmitter release, which depended on the presence of phosphorylatable Liprin-α3. Our findings indicate that PKC-mediated phosphorylation of Liprin-α3 triggers its phase separation and modulates active zone structure and function. |
format | Online Article Text |
id | pubmed-8144191 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-81441912021-06-07 PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure Emperador-Melero, Javier Wong, Man Yan Wang, Shan Shan H. de Nola, Giovanni Nyitrai, Hajnalka Kirchhausen, Tom Kaeser, Pascal S. Nat Commun Article The active zone of a presynaptic nerve terminal defines sites for neurotransmitter release. Its protein machinery may be organized through liquid–liquid phase separation, a mechanism for the formation of membrane-less subcellular compartments. Here, we show that the active zone protein Liprin-α3 rapidly and reversibly undergoes phase separation in transfected HEK293T cells. Condensate formation is triggered by Liprin-α3 PKC-phosphorylation at serine-760, and RIM and Munc13 are co-recruited into membrane-attached condensates. Phospho-specific antibodies establish phosphorylation of Liprin-α3 serine-760 in transfected cells and mouse brain tissue. In primary hippocampal neurons of newly generated Liprin-α2/α3 double knockout mice, synaptic levels of RIM and Munc13 are reduced and the pool of releasable vesicles is decreased. Re-expression of Liprin-α3 restored these presynaptic defects, while mutating the Liprin-α3 phosphorylation site to abolish phase condensation prevented this rescue. Finally, PKC activation in these neurons acutely increased RIM, Munc13 and neurotransmitter release, which depended on the presence of phosphorylatable Liprin-α3. Our findings indicate that PKC-mediated phosphorylation of Liprin-α3 triggers its phase separation and modulates active zone structure and function. Nature Publishing Group UK 2021-05-24 /pmc/articles/PMC8144191/ /pubmed/34031393 http://dx.doi.org/10.1038/s41467-021-23116-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Emperador-Melero, Javier Wong, Man Yan Wang, Shan Shan H. de Nola, Giovanni Nyitrai, Hajnalka Kirchhausen, Tom Kaeser, Pascal S. PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title | PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title_full | PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title_fullStr | PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title_full_unstemmed | PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title_short | PKC-phosphorylation of Liprin-α3 triggers phase separation and controls presynaptic active zone structure |
title_sort | pkc-phosphorylation of liprin-α3 triggers phase separation and controls presynaptic active zone structure |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8144191/ https://www.ncbi.nlm.nih.gov/pubmed/34031393 http://dx.doi.org/10.1038/s41467-021-23116-w |
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