| 2008 |
ODA16 (DAW1 ortholog in Chlamydomonas) functions as a cargo-specific adaptor between IFT particles and outer row dynein for efficient dynein transport into flagella. ODA16 localization depends on IFT, and it directly interacts with IFT complex B subunit IFT46, demonstrated by yeast two-hybrid, in vitro pull-down, and co-immunoprecipitation from flagellar extracts. Dynein extracted from wild-type axonemes can rebind to oda16 axonemes in vitro, consistent with a role in transport rather than subunit preassembly or binding-site formation. |
Yeast two-hybrid, in vitro pull-down, co-immunoprecipitation, in vitro dynein rebinding assay, IFT-dependent localization analysis |
The Journal of cell biology |
High |
18852297
|
| 2017 |
Crystal structure of Chlamydomonas ODA16 revealed an 80-residue N-terminal domain and a C-terminal 8-bladed β-propeller domain; both are required for binding to the N-terminal 147 residues of IFT46 (Kd ~200 nM), while only the C-terminal β-propeller is required for interaction with ODAs. This structural mapping defined the architectural model for ODA16-mediated IFT of outer dynein arms. |
X-ray crystallography, pull-down assays, dissociation constant measurement, domain mutagenesis |
The Journal of biological chemistry |
High |
28298440
|
| 2017 |
The N-terminus of IFT46 is specifically required for intraflagellar transport of outer arm dynein and its cargo-adaptor ODA16 into flagella. A suppressor allele expressing only the IFT46 C-terminal 240 amino acids restores IFT-B stability and flagellar length but fails to import ODA16 or outer arm dynein, establishing that IFT46 N-terminus, ODA16, and outer arm dynein interact for IFT of dynein. |
Genetic suppressor analysis, transposon insertion characterization, flagellar protein analysis by immunofluorescence/western blot in Chlamydomonas mutants |
Molecular biology of the cell |
High |
28701346
|
| 2020 |
The crystal structure of human ODA16 (DAW1) shows a C-terminal 8-bladed β-propeller with high overall structural similarity to Chlamydomonas ODA16, but the N-terminal domain has no visible electron density. Notably, size exclusion chromatography and pull-down experiments failed to detect a direct interaction between human ODA16 and IFT46, suggesting that additional factors are required for ciliary import of ODAs in human cells. |
X-ray crystallography, size exclusion chromatography, pull-down experiments with recombinant human proteins |
Protein science : a publication of the Protein Society |
Medium |
32239748
|
| 2020 |
Planarian Smed-DAW1 (ortholog of DAW1) is required for motile cilia function in multiciliated epidermis and protonephridia. RNAi knockdown caused locomotion defects and edema without initial loss of cilia number or length, indicating DAW1 loss impairs cilia motility rather than ciliogenesis per se. Extended RNAi resulted in shorter epidermal cilia and fewer ciliated protonephridia, indicating a role in homeostatic maintenance of ciliated structures. |
Systemic RNAi knockdown in planarian Schmidtea mediterranea, phenotypic analysis (locomotion, edema), cilia morphology assessment |
Development, growth & differentiation |
Medium |
32359074
|
| 2022 |
Zebrafish Daw1 facilitates the timely onset of robust cilia motility during early development. daw1 mutants show markedly reduced cilia motility during early development that subsequently recovers toward wild-type levels; the early motility deficit leads to laterality defects and body axis curves that self-correct when motility recovers, while later cilia-dependent processes are less affected. |
Zebrafish daw1 mutant analysis, cilia motility imaging, left-right patterning phenotype assessment |
Development (Cambridge, England) |
Medium |
35708608
|
| 2022 |
Biallelic DAW1 loss-of-function in humans causes distal type 2 outer dynein arm assembly defects in axonemal respiratory cilia proteins, explaining reduced cilia-induced fluid flow. Pathogenic DAW1 missense variants display reduced protein stability. In zebrafish, daw1 mutants showed reduced cilia motility and left-right patterning defects rescued by wild-type but not mutant daw1 expression. In early mouse embryos, Daw1 expression is limited to distal motile ciliated cells of the node. |
Genomic variant identification, electron microscopy of ciliary ultrastructure, particle tracking velocimetry, zebrafish rescue experiments with wild-type vs. mutant daw1, mouse embryo expression analysis, protein stability assessment |
Genetics in medicine : official journal of the American College of Medical Genetics |
High |
36074124
|
| 2024 |
Active ARL3 GTPases in Trypanosoma brucei cilia bind ODA16 and dissociate it from the IFT complex, functioning as a cargo-unloading mechanism. Depletion of ARL3 stabilizes ODA16–IFT interaction, causing ODA16 accumulation in cilia and defects in axonemal assembly. Interactions between human DAW1 (HsDAW1) and ARL GTPases are conserved, and these interactions are altered in HsDAW1 disease variants. |
ARL3 depletion in T. brucei (RNAi), co-immunoprecipitation, ciliary protein localization analysis, conservation study with human DAW1 and ARL GTPases, disease variant functional testing |
Science advances |
Medium |
39231220
|
| 2025 |
In silico AlphaPulldown screening identified IDA3 and Arl3 as direct interactors of ODA16. Biochemical and biophysical assays showed that a conserved N-terminal motif in IFT46 binds one face of the ODA16 structure, while IDA3 and Arl3 bind the opposite face (C-terminal β-propeller), enabling them to dissociate ODA16 from IFT46 through an allosteric mechanism, thereby releasing ODA cargo from the IFT machinery. |
AlphaPulldown in silico screening, structural modeling, biochemical binding assays, biophysical assays on Chlamydomonas and human proteins |
The Journal of biological chemistry |
Medium |
39880089
|