Simultaneous measurement of smoothened entry into and exit from the primary cilium
| Autor: | Philip A. Beachy, Jynho Kim, Xiaoyan Zheng, James Kim, Navdar Sever, Elaine Y. C. Hsia |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Recombinant Fusion Proteins
lcsh:Medicine Biology Receptors G-Protein-Coupled Mice Signaling proteins Fluorescent protein Animals Cilia lcsh:Science Molecular Biology Multidisciplinary Cilium lcsh:R Biology and Life Sciences Cell Biology Smoothened Receptor Hedgehog signaling pathway Transport protein Cell biology Cell staining Luminescent Proteins Protein Transport Microscopy Fluorescence NIH 3T3 Cells lcsh:Q Signal transduction Smoothened Research Article Developmental Biology Signal Transduction |
| Zdroj: | PLoS ONE, Vol 9, Iss 8, p e104070 (2014) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Ciliary accumulation of signaling proteins must result from a rate of ciliary entry that exceeds ciliary exit, but approaches for distinguishing ciliary entry vs. exit are lacking. Using a photoconvertible fluorescent protein tag, we establish an assay that allows a separate but simultaneous examination of ciliary entry and exit of the Hedgehog signaling protein Smoothened in individual cells. We show that KAAD-cyclopamine selectively blocks entry, whereas ciliobrevin interferes initially with exit and eventually with both entry and exit of ciliary Smoothened. Our study provides an approach to understanding regulation of ciliary entry vs. exit of Hedgehog signaling components as well as other ciliary proteins. |
| Databáze: | OpenAIRE |
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