isoTarget: a genetic method for analyzing the functional diversity of splicing isoformsin vivo
| Autor: | Ruonan Li, Yujia Hu, Macy W. Veling, Sarah Pizzano, Hao Liu, Limin Yang, Zhao W, Bing Ye |
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| Rok vydání: | 2020 |
| Předmět: |
Gene isoform
0303 health sciences animal structures fungi Alternative splicing Biology Subcellular localization Cell biology 03 medical and health sciences DSCAM 0302 clinical medicine medicine.anatomical_structure Proteome RNA splicing medicine Axon Gene 030217 neurology & neurosurgery 030304 developmental biology |
| Popis: | SUMMARYProtein isoforms generated by alternative splicing contribute to proteome diversity. Due to the lack of effective techniques, isoform-specific functions, expression, localization, and signaling mechanisms of endogenous proteinsin vivoare unknown for most genes. Here we report a genetic method, termedisoTarget, for blocking the expression of a targeted isoform without affecting the other isoforms and for conditional tagging the targeted isoform for multi-level analyses in select cells. ApplyingisoTargetto two mutually exclusive isoforms ofDrosophilaDscam, Dscam[TM1] and [TM2], we found that endogenous Dscam[TM1] is localized in dendrites while Dscam[TM2] is in both dendrites and axons. We demonstrate that the difference in subcellular localization between Dscam[TM1] and [TM2], rather than any difference in biochemical properties, leads to the two isoforms’ differential contributions to dendrite and axon development. Moreover, withisoTarget, we discovered that the subcellular enrichment of functional partners results in a DLK/Wallenda-Dscam[TM2]-Dock signaling cascade specifically in axons.isoTargetis an effective technique for studying how alternative splicing enhances proteome complexity. |
| Databáze: | OpenAIRE |
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