Dataset Information

The protein complex of kinesin motor KIF1C and its hereditary spastic paraplegia associated mutant G102A

ABSTRACT: Regulated local translation, RNA transport and protein localization are critical for spatio-temporal gene expression in neurons. The localization of mRNA and subsequent local protein synthesis contribute to neuronal functions like synaptogenesis, synapse pruning, axon guidance, axonal regeneration and synaptic plasticity. Thus, mutations in motor proteins and subsequent cargo transport failure lead to motoneuron diseases. The kinesin family member 1 C has been shown to play a role in different cargo transport alongside the microtubule network, but the pathomechanisms behind KIF1C deficiency mediated motoneuron diseases has not been deciphered yet. Additionally, the exon junction complex has been suggested as a molecular link between splicing and cytoplasmic mRNA localization and local translation. Here we identified KIF1C as a EJC transporter in neuronal cells. We investigated the KIF1C proteome in differentiated SH-SY5Y cells and found an interaction of KIF1C with EJC components and other RNA-binding proteins in KIF1C overexpressing SH-SY5Y cells. The interaction could be validated via immunoprecipitation and an endogenous eIF4A3 co-immunoprecipitation. The co-localization of eIF4A3 and RBM8A with wildtpye KIF1C at protrusions of SH-SY5Ys further confirms the interaction. The mislocalization of eIF4A3 and RMB8A due to KIF1C mutation suggests a transport of the EJC by KIF1C. Furthermore, the interaction of KIF1C with PABPC1 and EJC components is RNA mediated. We additionally demonstrate via complex capture with KIF1C overexpressing HEK293 cells that KIF1C interacts with RNA itself. We therefore suggest the interaction of KIF1C not only with the EJC, but with a complex containing RNA and the EJC. Hence KIF1C is a transporter of mRNA and the EJC.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Sh-sy5y Cell, Cell Line Cell

DISEASE(S): Hereditary Spastic Paraplegia

SUBMITTER: Karsten Boldt

LAB HEAD: Dr. Karsten Boldt

PROVIDER: PXD037387 | Pride | 2023-12-28

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	HOR3091-S02-A-HA-ctrl.raw	Raw
	HOR3091-S06-A-Kif1c-WT.raw	Raw
	HOR3091-S10-A-Kif1c-Mut.raw	Raw
	HOR3132-S02-A-HA-contrl.raw	Raw
	HOR3132-S06-A-Kif1c-WT.raw	Raw

Items per page:

1 - 5 of 18

Publications

The kinesin motor KIF1C is a putative transporter of the exon junction complex in neuronal cells.

Nagel Maike M Noss Marvin M Xu Jishu J Horn Nicola N Ueffing Marius M Boldt Karsten K Schuele Rebecca R

RNA (New York, N.Y.) 20221031

Neurons critically depend on regulated RNA localization and tight control of spatio-temporal gene expression to maintain their morphological and functional integrity. Mutations in the kinesin motor protein gene KIF1C cause Hereditary Spastic Paraplegia, an autosomal recessive disease leading to predominant degeneration of the long axons of central motoneurons. In this study we aimed to gain insight into the molecular function of KIF1C and understand how KIF1C dysfunction contributes to motoneuro ...[more]

PMID: 36316088

Similar Datasets

Project description:Background: SH-SY5Y cells exhibit a neuronal phenotype when treated with all-trans retinoic acid (RA), but the molecular mechanism of activation in the signaling pathway mediated by phosphatidylinositol 3-kinase (PI3K) is not sufficiently understood. To shed new light on the mechanism, we comprehensively compared the gene expression profiles between SK-N-SH cells and two subtypes of SH-SY5Y cells (SH-SY5Y-A and SH-SY5Y-E), each of which showed a different phenotype during RA-mediated differentiation. Results: SH-SY5Y-A cells differentiated in the presence of RA, whereas RA-treated SH-SY5Y-E cells required additional treatment with brain-derived neurotrophic factor (BDNF) for full differentiation. In combination with perturbation using a PI3K inhibitor, LY294002, we identified 386 genes and categorized them into two clusters dependent on the PI3K signaling pathway during RA-mediated differentiation in SH-SY5Y-A cells. Transcriptional regulation of the gene cluster was greatly reduced in SK-N-SH cells or partially impaired in SH-SY5Y-E cells in coincidence with a defect in the neuronal phenotype of these cell lines. Additional stimulation with BDNF induced a set of neural genes which were down-regulated in RA-treated SH-SY5Y-E cells but were abundant in the differentiated SH-SY5Y-A cells. Conclusions: We identified the gene clusters controlled by PI3K- and TRKB-mediated signaling pathways during differentiation in two subtypes of SH-SY5Y cells. TRKB-mediated bypass pathway compensates for the impaired neural functions generated by defects in several signaling pathways including PI3K in SH-SY5Y-E cells. The expression profiling data are useful for further studies to elucidate the signal transduction-transcriptional network including PI3K and/or TRKB. Experiment Overall Design: Human neuroblastomas, SK-N-SH (HTB-11) and SH-SY5Y-A cells (CRL-2266) were obtained from the American Type Culture Collection (ATCC). We also obtained SH-SY5Y-E cells (EC94030304) from the European Collection of Cell Cultures (ECACC). Tissue culture cells were maintained in D-MEM/F12 1:1 mixture supplemented with 15% FBS (Fetal Bovine Serum) and 1% NEAA (Non-essential amino acid) in a 5% CO2 humidified incubator at 37oC. The culture medium was changed twice a week. For the RA-inducible experiment, random culture cells from two clone subtypes of SH-SY5Y and SK-N-SH were seeded in laminin coated culture dishes (BioCoat Laminin Cellware; BD Biosciences, Billerica, MA, USA) for 1 day and then transferred to a medium containing 10 μM of RA in the presence or the absence of LY294002 (10μM) for five days. For BDNF-induced sequential differentiation of the SH-SY5Y-E strain, cells were washed with D-MEM/F12 twice after five days in the presence of RA and then incubated with 50 ng/ml of BDNF in D-MEM/F12 without serum for three days.