Sudarshan Gadadhar / Gonzalo Alvarez Viar / Jan Niklas Hansen / An Gong / Aleksandr Kostarev / Côme Ialy-Radio / Sophie Leboucher / Marjorie Whitfield / Ahmed Ziyyat / Aminata Touré / Luis Alvarez / Gaia Pigino / Carsten Janke /
PubMed Abstract
Posttranslational modifications of the microtubule cytoskeleton have emerged as key regulators of cellular functions, and their perturbations have been linked to a growing number of human pathologies. ...Posttranslational modifications of the microtubule cytoskeleton have emerged as key regulators of cellular functions, and their perturbations have been linked to a growing number of human pathologies. Tubulin glycylation modifies microtubules specifically in cilia and flagella, but its functional and mechanistic roles remain unclear. In this study, we generated a mouse model entirely lacking tubulin glycylation. Male mice were subfertile owing to aberrant beat patterns of their sperm flagella, which impeded the straight swimming of sperm cells. Using cryo-electron tomography, we showed that lack of glycylation caused abnormal conformations of the dynein arms within sperm axonemes, providing the structural basis for the observed dysfunction. Our findings reveal the importance of microtubule glycylation for controlled flagellar beating, directional sperm swimming, and male fertility.
EMDB-11942: "Tubulin glycylation controls axonemal dynein activity, flagellar beat and male fertility": Pre-pre-power stroke conformation of the mouse sperm outer dynein arms (ODAs). Both beta- and gamma-heavy chains are in pre-power stroke conformation. Cryo-ET, classification and sub-tomogram averaging. Method: EM (subtomogram averaging) / Resolution: 24.7 Å
EMDB-11943: "Tubulin glycylation controls axonemal dynein activity, flagellar beat and male fertility": Pre-post-power stroke conformation of the mouse sperm outer dynein arms (ODAs), with gamma-heavy chain in pre-power stroke conformation and beta-heavy chain in post-power stroke conformation. Cryo-ET, classification and sub-tomogram averaging. Method: EM (subtomogram averaging) / Resolution: 35.7 Å
EMDB-11944: Post-pre-power stroke conformation of the mouse sperm outer dynein arms (ODAs), with gamma-heavy chain in post-power stroke conformation and beta-heavy chain in pre-power stroke conformation. Reconstruction obtained by sub-tomogram classification and averaging from cryo-electron tomograms of active flagella of mouse sperm cells. Method: EM (subtomogram averaging) / Resolution: 38.4 Å
EMDB-11945: Post-post-power stroke conformation of the mouse sperm outer dynein arms (ODAs), with both gamma-heavy chain and beta-heavy chain in post-power stroke conformation. Reconstruction obtained with sub-tomogram classification and averaging from cryo-electron tomograms of active flagella of mouse sperm cells. Method: EM (subtomogram averaging) / Resolution: 34.5 Å
EMDB-11946: Axonemal 96nm-repeat from active sperm flagella of wild type mouse. Reconstruction obtained with cryo-ET and sub-tomogram averaging. Method: EM (subtomogram averaging) / Resolution: 24.0 Å
EMDB-11947: Axonemal 96nm-repeat from active sperm flagella of glycylation deficient mouse (Ttll3-/-Ttll8-/- mutation). Reconstruction obtained with cryo-ET and sub-tomogram averaging. Method: EM (subtomogram averaging) / Resolution: 33.0 Å
EMDB-11948: Axonemal 96nm-repeat from active flagella of wild type mouse, showing inner dynein arms (IDAs) in pre-power stroke conformations. Reconstruction obtained with classification and averaging of sub-tomograms from cryo-electron tomograms of wild type active flagella. Method: EM (subtomogram averaging) / Resolution: 30.7 Å
EMDB-11949: Axonemal 96nm-repeat from active flagella of wild type mouse, showing inner dynein arms (IDAs) in post-power stroke conformations. Reconstruction obtained with classification and averaging of sub-tomograms from cryo-electron tomograms of wild type active flagella. Method: EM (subtomogram averaging) / Resolution: 33.2 Å
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Mus musculus (house mouse)
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Mus musculus (house mouse)