| 2002 |
Rootletin is the major structural component of the ciliary rootlet; it forms detergent-insoluble filaments via its coiled-coil tail domain, assembles into parallel in-register homodimers and higher-order polymers through the tail domain alone, and its globular head domain is required for targeting to the basal body and binding to kinesin light chain. In retinal photoreceptors, rootlets anchor ER membranes along their length. |
Recombinant protein expression, detergent-insolubility assay, domain deletion analysis, immunoelectron microscopy, monoclonal antibody epitope mapping |
The Journal of cell biology |
High |
12427867
|
| 2005 |
Rootletin forms striking fibers emanating from the proximal ends of centrioles (shown by immunoelectron microscopy), interacts with C-Nap1, is phosphorylated by Nek2 kinase, and is displaced from centrosomes at mitosis onset. siRNA-mediated depletion of rootletin causes centrosome splitting, establishing its role in centrosome cohesion. |
Immunoelectron microscopy, Co-immunoprecipitation (C-Nap1 interaction), in vitro kinase assay (Nek2 phosphorylation), siRNA knockdown with centrosome splitting readout, overexpression fiber formation assay |
The Journal of cell biology |
High |
16203858
|
| 2005 |
Rootletin interacts physically with C-Nap1 in vivo; they colocalize at basal bodies/centrioles and show coordinated cell-cycle-dependent centriole association. Rootletin fibers connect basal bodies in ciliated cells and span between centrioles in nonciliated cells (unlike C-Nap1 which is restricted to centriole ends). Expression of C-Nap1 fragments dissociates rootletin fibers from centrioles, causing centrosome separation in interphase. |
Co-immunoprecipitation, immunofluorescence co-localization, ultrastructural analysis, dominant-negative C-Nap1 fragment expression, overexpression phenotyping |
Molecular biology of the cell |
High |
16339073
|
| 2013 |
C. elegans CHE-10/rootletin ortholog maintains ciliary integrity by modulating assembly, motility, and flux of IFT particles (axoneme length control) and by stabilizing ciliary transition zones and basal bodies. Loss of CHE-10/rootletin disrupts periciliary membrane organization, impairing delivery of basal body-associated and ciliary components and causing cilium degeneration. |
Genetic mutant analysis (che-10), in vivo IFT particle tracking, fluorescence imaging of transition zones and basal bodies, epistasis with IFT mutants |
Current biology : CB |
High |
24094853
|
| 2015 |
Drosophila Rootletin assembles into rootlets of diverse lengths in sensory neurons, is required for basal body cohesion and sensory function (mechanosensation and chemosensation), and requires centrioles for normal rootlet assembly. The N-terminal conserved domain of Root is essential for its function in vivo. Rootletin localizes asymmetrically to mother centrosomes in neuroblasts and to mother centrioles in spermatocytes, both requiring Bld10. |
Drosophila genetic knockout, behavioral assays, immunofluorescence, domain deletion rescue experiments, Bld10 epistasis |
The Journal of cell biology |
Medium |
26483560
|
| 2015 |
Drosophila Rootletin knockdown results in loss of ciliary rootlet in chordotonal neurons and severe disruption of mechanosensory function, while cilium structure and protein localization appear largely normal, suggesting rootletin's role is in anchoring/mechanotransduction rather than ciliogenesis per se. |
RNAi knockdown in Drosophila, electron microscopy of cilia, electrophysiological/behavioral mechanosensory assays |
Cilia |
Medium |
26140210
|
| 2012 |
siRNA depletion of rootletin or C-NAP1 increases radiation-induced centriole splitting, and rootletin or C-NAP1 knockdown reduces primary cilium formation, establishing that the centriole cohesion apparatus at the proximal end of centrioles facilitates ciliogenesis. |
siRNA knockdown, gamma irradiation, immunofluorescence scoring of centriole splitting and primary cilia formation |
Cell cycle (Georgetown, Tex.) |
Medium |
23070519
|
| 2017 |
Rootletin prevents VHL E3 ligase-mediated proteasomal degradation of Cep68: in the absence of rootletin, VHL ubiquitinates Cep68 both in vitro and in vivo, leading to Cep68 loss and centrosome splitting. Co-silencing of rootletin and VHL rescues Cep68 levels and centrosome cohesion. A Cep68 mutant that cannot bind the VHL β-domain also suppresses centrosome splitting caused by rootletin depletion. |
In vitro ubiquitination assay, siRNA knockdown of rootletin and VHL (single and double), Cep68 ubiquitination-resistant mutants, immunofluorescence centrosome cohesion assay |
Biochimica et biophysica acta. Molecular cell research |
High |
28089774
|
| 2018 |
STED nanoscopy revealed that C-Nap1 forms a ring at each centriole's proximal end that organizes a rootletin ring and multiple rootletin/CEP68 fibers. Rootletin/CEP68 fibers from the two centrosomes form an interdigitating network. Rootletin molecules within filaments are staggered N-to-N and C-to-C at 75-nm intervals; rootletin binds CEP68 via its C-terminal spectrin repeat-containing region at 75-nm intervals; estimated N-to-C rootletin length is ~35–40 nm, minimal rootletin length ~110 nm. CEP68 is required to form rootletin filaments branching from centrioles and modulates fiber thickness. |
STED super-resolution microscopy, protein interaction mapping (binding domain identification), siRNA depletion of CEP68 with filament phenotype readout |
Proceedings of the National Academy of Sciences of the United States of America |
High |
29463719
|
| 2020 |
Cep44 localizes to the proximal end of centrioles, associates with rootletin, and regulates rootletin's stability and localization to the centrosome. Cep44 ablation leads to loss of centrosome cohesion; this is independent of C-Nap1, LRRC45, and Cep215 stability/recruitment. |
siRNA knockdown, co-immunoprecipitation (Cep44–rootletin interaction), immunofluorescence localization, protein stability assays |
Journal of cell science |
Medium |
31974111
|
| 2021 |
Rootletin, ELMOD2, and ARL2 act in a common pathway to suppress spurious ciliation and maintain centrosome cohesion. Rootletin deletion phenotypes (increased ciliation, multiciliation, loss of centrosome cohesion) are rescued by increasing ARL2 activity but not ELMOD2 overexpression. Epistasis analysis places this pathway downstream of TTBK2 and upstream of CP110, preventing spurious CP110 release and regulating ciliary vesicle docking. |
Mouse embryonic fibroblast genetic deletion (Rootletin KO), ARL2 overexpression rescue, epistasis with TTBK2/CP110 markers, immunofluorescence for ciliation and cohesion markers |
Molecular biology of the cell |
Medium |
33596093
|
| 2024 |
Rootletin molecules in mouse hippocampal neurons exhibit highly organized intracellular distributions, and formation of ciliary rootlets precedes that of primary cilia, as revealed by label-free second harmonic generation imaging. |
Second harmonic generation microscopy combined with coherent anti-Stokes Raman scattering imaging in mouse brain tissue (label-free) |
bioRxivpreprint |
Low |
bio_10.1101_2024.06.19.597702
|