The role of the dynein light intermediate chain in retrograde IFT and flagellar function inChlamydomonas
| Autor: | Alexandria Schauer, Catherine A. Perrone, Raqual Bower, Kristyn Van Der Waal Mills, Mary E. Porter, Douglas Tritschler, Jaimee Reck |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Dynein Motility Flagellum Microtubules 03 medical and health sciences Microtubule Intraflagellar transport Animals Cilia Molecular Biology Cytoskeleton biology Cilium Chlamydomonas Dyneins Articles Cell Biology biology.organism_classification 3. Good health Transport protein Cell biology Protein Transport 030104 developmental biology Flagella sense organs |
| Zdroj: | Molecular Biology of the Cell |
| ISSN: | 1939-4586 1059-1524 |
| Popis: | D1bLIC is a subunit of the retrograde IFT motor. Knockdown or knockout of D1bLIC has dose-dependent effects on flagellar assembly, length, motility, and signaling. iTRAQ-based proteomics identifies novel proteins altered in d1blic mutant flagella. TIRF microscopy reveals the kinetics and remodeling of the retrograde motor at the flagellar tip. The assembly of cilia and flagella depends on the activity of two microtubule motor complexes, kinesin-2 and dynein-2/1b, but the specific functions of the different subunits are poorly defined. Here we analyze Chlamydomonas strains expressing different amounts of the dynein 1b light intermediate chain (D1bLIC). Disruption of D1bLIC alters the stability of the dynein 1b complex and reduces both the frequency and velocity of retrograde intraflagellar transport (IFT), but it does not eliminate retrograde IFT. Flagellar assembly, motility, gliding, and mating are altered in a dose-dependent manner. iTRAQ-based proteomics identifies a small subset of proteins that are significantly reduced or elevated in d1blic flagella. Transformation with D1bLIC-GFP rescues the mutant phenotypes, and D1bLIC-GFP assembles into the dynein 1b complex at wild-type levels. D1bLIC-GFP is transported with anterograde IFT particles to the flagellar tip, dissociates into smaller particles, and begins processive retrograde IFT in |
| Databáze: | OpenAIRE |
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