Resident CAPS on dense-core vesicles docks and primes vesicles for fusion
The Ca[superscript 2+]-dependent exocytosis of dense-core vesicles in neuroendocrine cells requires a priming step during which SNARE protein complexes assemble. CAPS (aka CADPS) is one of several factors required for vesicle priming; however, the localization and dynamics of CAPS at sites of exocyt...
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| Формат: | Стаття |
| Мова: | en_US |
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American Society for Cell Biology
2016
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| Онлайн доступ: | http://hdl.handle.net/1721.1/102616 https://orcid.org/0000-0003-4363-441X |
| _version_ | 1826194709172715520 |
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| author | Kielar-Grevstad, D. Michelle Zhang, Xingmin James, Declan J. Martin, Thomas F. J. Kabachinski, Gregory L. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Kielar-Grevstad, D. Michelle Zhang, Xingmin James, Declan J. Martin, Thomas F. J. Kabachinski, Gregory L. |
| author_sort | Kielar-Grevstad, D. Michelle |
| collection | MIT |
| description | The Ca[superscript 2+]-dependent exocytosis of dense-core vesicles in neuroendocrine cells requires a priming step during which SNARE protein complexes assemble. CAPS (aka CADPS) is one of several factors required for vesicle priming; however, the localization and dynamics of CAPS at sites of exocytosis in live neuroendocrine cells has not been determined. We imaged CAPS before, during, and after single-vesicle fusion events in PC12 cells by TIRF microscopy. In addition to being a resident on cytoplasmic dense-core vesicles, CAPS was present in clusters of approximately nine molecules near the plasma membrane that corresponded to docked/tethered vesicles. CAPS accompanied vesicles to the plasma membrane and was present at all vesicle exocytic events. The knockdown of CAPS by shRNA eliminated the VAMP-2–dependent docking and evoked exocytosis of fusion-competent vesicles. A CAPS(ΔC135) protein that does not localize to vesicles failed to rescue vesicle docking and evoked exocytosis in CAPS-depleted cells, showing that CAPS residence on vesicles is essential. Our results indicate that dense-core vesicles carry CAPS to sites of exocytosis, where CAPS promotes vesicle docking and fusion competence, probably by initiating SNARE complex assembly. |
| first_indexed | 2024-09-23T10:00:33Z |
| format | Article |
| id | mit-1721.1/102616 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:00:33Z |
| publishDate | 2016 |
| publisher | American Society for Cell Biology |
| record_format | dspace |
| spelling | mit-1721.1/1026162022-09-26T15:09:51Z Resident CAPS on dense-core vesicles docks and primes vesicles for fusion Kielar-Grevstad, D. Michelle Zhang, Xingmin James, Declan J. Martin, Thomas F. J. Kabachinski, Gregory L. Massachusetts Institute of Technology. Department of Biology Kabachinski, Greg The Ca[superscript 2+]-dependent exocytosis of dense-core vesicles in neuroendocrine cells requires a priming step during which SNARE protein complexes assemble. CAPS (aka CADPS) is one of several factors required for vesicle priming; however, the localization and dynamics of CAPS at sites of exocytosis in live neuroendocrine cells has not been determined. We imaged CAPS before, during, and after single-vesicle fusion events in PC12 cells by TIRF microscopy. In addition to being a resident on cytoplasmic dense-core vesicles, CAPS was present in clusters of approximately nine molecules near the plasma membrane that corresponded to docked/tethered vesicles. CAPS accompanied vesicles to the plasma membrane and was present at all vesicle exocytic events. The knockdown of CAPS by shRNA eliminated the VAMP-2–dependent docking and evoked exocytosis of fusion-competent vesicles. A CAPS(ΔC135) protein that does not localize to vesicles failed to rescue vesicle docking and evoked exocytosis in CAPS-depleted cells, showing that CAPS residence on vesicles is essential. Our results indicate that dense-core vesicles carry CAPS to sites of exocytosis, where CAPS promotes vesicle docking and fusion competence, probably by initiating SNARE complex assembly. 2016-05-23T15:22:20Z 2016-05-23T15:22:20Z 2015-12 2015-12 Article http://purl.org/eprint/type/JournalArticle 1059-1524 1939-4586 http://hdl.handle.net/1721.1/102616 Kabachinski, G., D. M. Kielar-Grevstad, X. Zhang, D. J. James, and T. F. J. Martin. “Resident CAPS on Dense-Core Vesicles Docks and Primes Vesicles for Fusion.” Molecular Biology of the Cell 27, no. 4 (February 15, 2016): 654–68. https://orcid.org/0000-0003-4363-441X en_US http://dx.doi.org/10.1091/mbc.E15-07-0509 Molecular Biology of the Cell Creative Commons Attribution http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf American Society for Cell Biology American Society for Cell Biology |
| spellingShingle | Kielar-Grevstad, D. Michelle Zhang, Xingmin James, Declan J. Martin, Thomas F. J. Kabachinski, Gregory L. Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title | Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title_full | Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title_fullStr | Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title_full_unstemmed | Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title_short | Resident CAPS on dense-core vesicles docks and primes vesicles for fusion |
| title_sort | resident caps on dense core vesicles docks and primes vesicles for fusion |
| url | http://hdl.handle.net/1721.1/102616 https://orcid.org/0000-0003-4363-441X |
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