STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena
Abstract The base of the cilium comprising the transition zone (TZ) and transition fibers (TF) acts as a selecting gate to regulate the intraflagellar transport (IFT)-dependent trafficking of proteins to and from cilia. Before entering the ciliary compartment, IFT complexes and transported cargoes a...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-86909-5 |
| _version_ | 1819039973731467264 |
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| author | Khodor S. Hazime Zhu Zhou Ewa Joachimiak Natalia A. Bulgakova Dorota Wloga Jarema J. Malicki |
| author_facet | Khodor S. Hazime Zhu Zhou Ewa Joachimiak Natalia A. Bulgakova Dorota Wloga Jarema J. Malicki |
| author_sort | Khodor S. Hazime |
| collection | DOAJ |
| description | Abstract The base of the cilium comprising the transition zone (TZ) and transition fibers (TF) acts as a selecting gate to regulate the intraflagellar transport (IFT)-dependent trafficking of proteins to and from cilia. Before entering the ciliary compartment, IFT complexes and transported cargoes accumulate at or near the base of the cilium. The spatial organization of IFT proteins at the cilia base is key for understanding cilia formation and function. Using stochastic optical reconstruction microscopy (STORM) and computational averaging, we show that seven TZ, nine IFT, three Bardet–Biedl syndrome (BBS), and one centrosomal protein, form 9-clustered rings at the cilium base of a ciliate Tetrahymena thermophila. In the axial dimension, analyzed TZ proteins localize to a narrow region of about 30 nm while IFT proteins dock approximately 80 nm proximal to TZ. Moreover, the IFT-A subcomplex is positioned peripheral to the IFT-B subcomplex and the investigated BBS proteins localize near the ciliary membrane. The positioning of the HA-tagged N- and C-termini of the selected proteins enabled the prediction of the spatial orientation of protein particles and likely cargo interaction sites. Based on the obtained data, we built a comprehensive 3D-model showing the arrangement of the investigated ciliary proteins. |
| first_indexed | 2024-12-21T09:01:43Z |
| format | Article |
| id | doaj.art-ad03647e47844e35a88866b74e826023 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-21T09:01:43Z |
| publishDate | 2021-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-ad03647e47844e35a88866b74e8260232022-12-21T19:09:28ZengNature PortfolioScientific Reports2045-23222021-04-0111111510.1038/s41598-021-86909-5STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in TetrahymenaKhodor S. Hazime0Zhu Zhou1Ewa Joachimiak2Natalia A. Bulgakova3Dorota Wloga4Jarema J. Malicki5Bateson Centre and the Department of Biomedical Science, University of SheffieldBateson Centre and the Department of Biomedical Science, University of SheffieldLaboratory of Cytoskeleton and Cilia BiologyBateson Centre and the Department of Biomedical Science, University of SheffieldLaboratory of Cytoskeleton and Cilia BiologyBateson Centre and the Department of Biomedical Science, University of SheffieldAbstract The base of the cilium comprising the transition zone (TZ) and transition fibers (TF) acts as a selecting gate to regulate the intraflagellar transport (IFT)-dependent trafficking of proteins to and from cilia. Before entering the ciliary compartment, IFT complexes and transported cargoes accumulate at or near the base of the cilium. The spatial organization of IFT proteins at the cilia base is key for understanding cilia formation and function. Using stochastic optical reconstruction microscopy (STORM) and computational averaging, we show that seven TZ, nine IFT, three Bardet–Biedl syndrome (BBS), and one centrosomal protein, form 9-clustered rings at the cilium base of a ciliate Tetrahymena thermophila. In the axial dimension, analyzed TZ proteins localize to a narrow region of about 30 nm while IFT proteins dock approximately 80 nm proximal to TZ. Moreover, the IFT-A subcomplex is positioned peripheral to the IFT-B subcomplex and the investigated BBS proteins localize near the ciliary membrane. The positioning of the HA-tagged N- and C-termini of the selected proteins enabled the prediction of the spatial orientation of protein particles and likely cargo interaction sites. Based on the obtained data, we built a comprehensive 3D-model showing the arrangement of the investigated ciliary proteins.https://doi.org/10.1038/s41598-021-86909-5 |
| spellingShingle | Khodor S. Hazime Zhu Zhou Ewa Joachimiak Natalia A. Bulgakova Dorota Wloga Jarema J. Malicki STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena Scientific Reports |
| title | STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena |
| title_full | STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena |
| title_fullStr | STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena |
| title_full_unstemmed | STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena |
| title_short | STORM imaging reveals the spatial arrangement of transition zone components and IFT particles at the ciliary base in Tetrahymena |
| title_sort | storm imaging reveals the spatial arrangement of transition zone components and ift particles at the ciliary base in tetrahymena |
| url | https://doi.org/10.1038/s41598-021-86909-5 |
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