Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication
Duplication of the yeast centrosome (called the spindle pole body, SPB) is thought to occur through a series of discrete steps that culminate in insertion of the new SPB into the nuclear envelope (NE). To better understand this process, we developed a novel two-color structured illumination microsco...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2015-09-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/08586 |
| _version_ | 1818018866625773568 |
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| author | Shannon Burns Jennifer S Avena Jay R Unruh Zulin Yu Sarah E Smith Brian D Slaughter Mark Winey Sue L Jaspersen |
| author_facet | Shannon Burns Jennifer S Avena Jay R Unruh Zulin Yu Sarah E Smith Brian D Slaughter Mark Winey Sue L Jaspersen |
| author_sort | Shannon Burns |
| collection | DOAJ |
| description | Duplication of the yeast centrosome (called the spindle pole body, SPB) is thought to occur through a series of discrete steps that culminate in insertion of the new SPB into the nuclear envelope (NE). To better understand this process, we developed a novel two-color structured illumination microscopy with single-particle averaging (SPA-SIM) approach to study the localization of all 18 SPB components during duplication using endogenously expressed fluorescent protein derivatives. The increased resolution and quantitative intensity information obtained using this method allowed us to demonstrate that SPB duplication begins by formation of an asymmetric Sfi1 filament at mitotic exit followed by Mps1-dependent assembly of a Spc29- and Spc42-dependent complex at its tip. Our observation that proteins involved in membrane insertion, such as Mps2, Bbp1, and Ndc1, also accumulate at the new SPB early in duplication suggests that SPB assembly and NE insertion are coupled events during SPB formation in wild-type cells. |
| first_indexed | 2024-04-14T07:45:19Z |
| format | Article |
| id | doaj.art-8d1c0ba962874327a7dd1545b85446f1 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-14T07:45:19Z |
| publishDate | 2015-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-8d1c0ba962874327a7dd1545b85446f12022-12-22T02:05:21ZengeLife Sciences Publications LtdeLife2050-084X2015-09-01410.7554/eLife.08586Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplicationShannon Burns0Jennifer S Avena1Jay R Unruh2Zulin Yu3Sarah E Smith4Brian D Slaughter5Mark Winey6Sue L Jaspersen7Stowers Institute for Medical Research, Kansas City, United StatesDepartment of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United StatesStowers Institute for Medical Research, Kansas City, United StatesStowers Institute for Medical Research, Kansas City, United StatesStowers Institute for Medical Research, Kansas City, United StatesStowers Institute for Medical Research, Kansas City, United StatesDepartment of Molecular, Cellular and Developmental Biology, University of Colorado Boulder, Boulder, United StatesStowers Institute for Medical Research, Kansas City, United States; Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, United StatesDuplication of the yeast centrosome (called the spindle pole body, SPB) is thought to occur through a series of discrete steps that culminate in insertion of the new SPB into the nuclear envelope (NE). To better understand this process, we developed a novel two-color structured illumination microscopy with single-particle averaging (SPA-SIM) approach to study the localization of all 18 SPB components during duplication using endogenously expressed fluorescent protein derivatives. The increased resolution and quantitative intensity information obtained using this method allowed us to demonstrate that SPB duplication begins by formation of an asymmetric Sfi1 filament at mitotic exit followed by Mps1-dependent assembly of a Spc29- and Spc42-dependent complex at its tip. Our observation that proteins involved in membrane insertion, such as Mps2, Bbp1, and Ndc1, also accumulate at the new SPB early in duplication suggests that SPB assembly and NE insertion are coupled events during SPB formation in wild-type cells.https://elifesciences.org/articles/08586spindle pole bodycentrosomestructured illumination microscopySfi1Cdc31/centrinsingle particle averaging |
| spellingShingle | Shannon Burns Jennifer S Avena Jay R Unruh Zulin Yu Sarah E Smith Brian D Slaughter Mark Winey Sue L Jaspersen Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication eLife spindle pole body centrosome structured illumination microscopy Sfi1 Cdc31/centrin single particle averaging |
| title | Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| title_full | Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| title_fullStr | Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| title_full_unstemmed | Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| title_short | Structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| title_sort | structured illumination with particle averaging reveals novel roles for yeast centrosome components during duplication |
| topic | spindle pole body centrosome structured illumination microscopy Sfi1 Cdc31/centrin single particle averaging |
| url | https://elifesciences.org/articles/08586 |
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