Basal bodies (BBs) are cellular nanomachines that position and anchor cilia. Defects in BBs contribute to a broad spectrum of diseases including cancers and ciliopathies. Ciliopathies are a general class of human maladies that include birth defects, polydactyly, blindness, respiratory illness, hydrocephaly, and infertility. However, the molecular mechanics for how BBs organize cilia so that, when defective, they contribute to the above pathologies are not well understood. Motile cilia produce large-scale hydrodynamic fluid flow in the respiratory tract, brain ventricles and oviduct. Loss of directional fluid flow causes respiratory illness, hydrocephalus and infertility. While study of such human diseases reveals the importance of BBs and cilia, fundamental cell biology research into the mechanisms by which BBs position and anchor cilia to promote normal ciliary beating is in part limited due to a need for genetically modifiable model systems in which human disease mutations can be mimicked and where ciliary beating is accessible. Here we build on our prior funding cycle where we developed Tetrahymena as a model system to visualize ciliary forces and identified proteins and post-translational modifications responsible for normal ciliary positioning and anchoring. Our proposed funding will capitalize on these prior advances to quantify the forces that asymmetric cilia beating imposes on BBs. Such forces are predicted from computational modeling, but the direct BB movements that arise from these forces have never been studied. To ensure that BBs and cilia are correctly positioned within ciliary arrays, BBs possess BB-associated accessory structures. We will determine how these accessory structures dynamically respond to ciliary forces to preserve BB and ciliary structure and positioning to maintain proper hydrodynamic flow. Next, we will study the BB domains that ensure proper positioning and anchorage at the cell surface during ciliary beating. Ciliary forces induce asymmetric disruption of specific triplet microtubules in BB mutants. We will establish how asymmetrically localized BB stability factors define distinct BB domains and determine whether they stabilize particular BB triplet microtubules. These studies will be carried out by a collaborative group of researchers that bridges a broad research spectrum from fundamental biology, genetics, live and fixed cell microscopy, polymer engineering, computational modeling to structural studies. Excellent training opportunities exist for undergraduate and graduate students and postdoctoral researchers in the lab. Collaborations with other labs that specialize in light and electron microscopy, computational modeling, structural biology and polymer engineering, expand the innovation and impact of our studies. In summary, this collaborative proposal will illuminate how BBs position cilia and withstand the forces generated by cilia so that coherent hydrodynamic forces are generated and maintained.
Basal bodies are essential for the formation of cilia, cellular appendages that enable cells to sense their environment and to produce fluid flow in the body. Loss of cilia leads to embryonic lethality and ciliary defects contribute to a devastating class of diseases with symptoms that can include respiratory illness, hydrocephalus, infertility, blindness, obesity, cancer, neurological impairment, and polycystic kidneys. Our research aims to discover the etiology of such maladies.
|Louka, Panagiota; Vasudevan, Krishna Kumar; Guha, Mayukh et al. (2018) Proteins that control the geometry of microtubules at the ends of cilia. J Cell Biol 217:4298-4313|
|Galati, Domenico F; Sullivan, Kelly D; Pham, Andrew T et al. (2018) Trisomy 21 Represses Cilia Formation and Function. Dev Cell 46:641-650.e6|
|Baschal, Erin E; Terhune, Elizabeth A; Wethey, Cambria I et al. (2018) Idiopathic Scoliosis Families Highlight Actin-Based and Microtubule-Based Cellular Projections and Extracellular Matrix in Disease Etiology. G3 (Bethesda) 8:2663-2672|
|Kendrick, Agnieszka A; Schafer, Johnathon; Dzieciatkowska, Monika et al. (2017) CD147: a small molecule transporter ancillary protein at the crossroad of multiple hallmarks of cancer and metabolic reprogramming. Oncotarget 8:6742-6762|
|Wall, Kathryn P; Pagratis, Maria; Armstrong, Geoffrey et al. (2016) Molecular Determinants of Tubulin's C-Terminal Tail Conformational Ensemble. ACS Chem Biol 11:2981-2990|
|Ruehle, Marisa D; Orias, Eduardo; Pearson, Chad G (2016) Tetrahymena as a Unicellular Model Eukaryote: Genetic and Genomic Tools. Genetics 203:649-65|
|Galati, Domenico F; Mitchell, Brian J; Pearson, Chad G (2016) Subdistal Appendages Stabilize the Ups and Downs of Ciliary Life. Dev Cell 39:387-389|
|Bayless, Brian A; Galati, Domenico F; Junker, Anthony D et al. (2016) Asymmetrically localized proteins stabilize basal bodies against ciliary beating forces. J Cell Biol 215:457-466|
|Meehl, Janet B; Bayless, Brian A; Giddings Jr, Thomas H et al. (2016) Tetrahymena Poc1 ensures proper intertriplet microtubule linkages to maintain basal body integrity. Mol Biol Cell 27:2394-403|
|Galati, Domenico F (2016) Pkd Proteins Team Up to Tell Cilia Which Way to Go. J Neurosci 36:643-5|
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