Keywords: KIF1C, Ataxia, Myelin, oligodendrocytes Internship Duration: 30/11/-1 - 30/11/-1
Head of the hosting team: Hervé Acloque - Katayoun Moazami-Goudarzi
Address of the host laboratory: Génétique Animale et Biologie Intégrative Team Etudes Fonctionnelles et Modèles Innovants pour la Santé des Animaux d'Élevage Domaine de Vilvert Bat440 78350 Jouy en Josas France
Supervisor: Amandine DuchesneE-mail: amandine.duchesne@inrae.fr Phone: 0134652672
Context : KIF1C is a widely expressed kinesin-3 motor protein, which transports a variety of cargoes, such as dense-core vesicles into axons and dendrites, alpha5beta1 integrins in migrating cells or APC-dependent mRNAs in cell protrusions. Thus, KIF1C functions are likely to be highly context-specific and its biological role(s) may vary from one cell type to another. KIF1C mutations are associated in human with a rare form of spastic ataxia (SPAX2/SPG58; Caballero Oteyza et al. 2014 ; Dor et al., 2014), characterized by progressive spastic paraplegia and cerebellar ataxia. KIF1C loss-of-function is also involved in a frequent recessive form of demyelinating ataxia in Charolais cattle (Duchesne et al., 2018) associated with dysfunctional oligodendrocytes, highlighting a new role of KIF1C in preserving the structural integrity and function of myelin. We recently established the first mouse model mimicking the bovine KIF1C loss-of-function mutation in a pure genetic background. Basic locomotion tests were concordant with a late onset ataxic gait. Neuropathological analysis revealed early and progressive abnormal myelin structures. Transcriptomic analyses showed that KIF1C deficiency has an early impact on myelination with a downregulation of genes involved in gliogenesis and myelination. Moreover, some of these downregulated genes were also mislocalized, supporting the hypothesis of KIF1C being involved in the transport of myelin protein mRNAs in oligodendrocytes, either directly or in interaction with ribonucleoprotein (RNP) granules. Objectives: In this project, the candidate will identify KIF1C-cargoes in oligodendrocyte primary cultures. To this aim, he/she will take advantage of an in-lab established mouse model with a V5-tagged KIF1C protein (unpublished data). This model will facilitate and enhance specificity of KIF1C affinity purification. He/she will first derive oligodendrocyte primary cultures from the KIF1C V5-tagged mouse model. Then, he/she will identify proteins and RNAs interacting with KIF1C in oligodendrocyte primary cultures using immunoprecipitation, MS-MS / RNA-immunoprecipitation and RNAseq analyses.
Isolation and culture of oligodendrocytes Immunoprecipitation / RNA immunoprecipitation MSMS and RNAseq analyses
Duchesne et al., Progressive ataxia of Charolais cattle highlights a role of KIF1C in sustainable myelination. PLoS Genet. 2018 Aug 1;14(8):e1007550. doi: 10.1371/journal.pgen.1007550. Caballero Oteyza et al., Motor protein mutations cause a new form of hereditary spastic paraplegia. Neurology. 2014 Jun 3;82(22):2007-16. doi: 10.1212/WNL.0000000000000479. Pichon et al., The kinesin KIF1C transports APC-dependent mRNAs to cell protrusions. RNA. 2021 Dec;27(12):1528-1544. doi: 10.1261/rna.078576.120.
