Fast Transport of RNA Granules by Direct Interactions with KIF5A/KLC1 Motors Prevents Axon Degeneration

Complex neural circuitry requires stable connections formed by lengthy axons. To maintain these functional circuits, fast transport delivers RNAs to distal axons where they undergo local translation. However, the mechanism that enables long distance transport of non-membrane enclosed organelles such as RNA granules is not known. Here we demonstrate that a complex containing RNA and the RNA-binding protein (RBP) SFPQ interacts directly with a tetrameric kinesin containing the adaptor KLC1 and the motor KIF5A. We show that binding of SFPQ to KIF5A/KLC1 motor complex is required for axon survival and is impacted by KIF5A mutations that cause Charcot-Marie-Tooth (CMT) Disease. Moreover, therapeutic approaches that bypass the need for local translation of SFPQ-bound proteins prevent axon degeneration in CMT models. Collectively, these observations show that non-membrane enclosed organelles can move autonomously and that replacing axonally translated proteins provides a therapeutic approach to axonal degenerative disorders.

[1]  Michael S. Fernandopulle,et al.  RNA Granules Hitchhike on Lysosomes for Long-Distance Transport, Using Annexin A11 as a Molecular Tether , 2019, Cell.

[2]  Anne E Carpenter,et al.  A High-Content Screen Identifies TPP1 and Aurora B as Regulators of Axonal Mitochondrial Transport , 2019, Cell reports.

[3]  G. Schiavo,et al.  Axonal transport and neurological disease , 2019, Nature Reviews Neurology.

[4]  R. Singer,et al.  The travels of mRNAs in neurons: do they know where they are going? , 2019, Current Opinion in Neurobiology.

[5]  S. Caria,et al.  A new crystal structure and small-angle X-ray scattering analysis of the homodimer of human SFPQ. , 2019, Acta crystallographica. Section F, Structural biology communications.

[6]  J. Onuchic,et al.  Structural consequences of hereditary spastic paraplegia disease-related mutations in kinesin , 2018, Proceedings of the National Academy of Sciences.

[7]  M. Aumont-Nicaise,et al.  Characterization of the binding mode of JNK-interacting protein 1 (JIP1) to kinesin-light chain 1 (KLC1) , 2018, The Journal of Biological Chemistry.

[8]  C. Bond,et al.  Functional Domains of NEAT1 Architectural lncRNA Induce Paraspeckle Assembly through Phase Separation. , 2018, Molecular cell.

[9]  J. Ule,et al.  Intron retention and nuclear loss of SFPQ are molecular hallmarks of ALS , 2018, Nature Communications.

[10]  R. Steiner,et al.  Structural basis for isoform-specific kinesin-1 recognition of Y-acidic cargo adaptors , 2018, bioRxiv.

[11]  Timothy A. Miller,et al.  Genome-wide Analyses Identify KIF5A as a Novel ALS Gene in and for Therapeutic , 2018 .

[12]  R. Segal,et al.  Paclitaxel Reduces Axonal Bclw to Initiate IP3R1-Dependent Axon Degeneration , 2017, Neuron.

[13]  William M Alexander,et al.  multiplierz v2.0: A Python‐based ecosystem for shared access and analysis of native mass spectrometry data , 2017, Proteomics.

[14]  Madhusoodanan Mottamal,et al.  Characterization of kinesin switch I mutations that cause hereditary spastic paraplegia , 2017, PloS one.

[15]  W. Talbot,et al.  Non-nuclear Pool of Splicing Factor SFPQ Regulates Axonal Transcripts Required for Normal Motor Development , 2017, Neuron.

[16]  Jeffrey J. Nirschl,et al.  Amyotrophic lateral sclerosis-linked mutations increase the viscosity of liquid-like TDP-43 RNP granules in neurons , 2017, Proceedings of the National Academy of Sciences.

[17]  Byung-Ok Choi,et al.  Wide phenotypic spectrum in axonal Charcot–Marie–Tooth neuropathy type 2 patients with KIF5A mutations , 2017, Genes & Genomics.

[18]  E. Betzig,et al.  Engulfed cadherin fingers are polarized junctional structures between collectively migrating endothelial cells , 2016, Nature Cell Biology.

[19]  Hunter L. Elliott,et al.  The RNA-binding protein SFPQ orchestrates an RNA regulon to promote axon viability , 2016, Nature Neuroscience.

[20]  C. Hoogenraad,et al.  Microtubule‐binding protein doublecortin‐like kinase 1 (DCLK1) guides kinesin‐3‐mediated cargo transport to dendrites , 2016, The EMBO journal.

[21]  L. Walensky,et al.  Inhibition of Pro-apoptotic BAX by a noncanonical interaction mechanism. , 2015, Molecular cell.

[22]  R. Segal,et al.  Campenot Cultures and Microfluidics Provide Complementary Platforms for Spatial Study of Dorsal Root Ganglia Neurons , 2015 .

[23]  D. Hughes,et al.  Extended phenotypic spectrum of KIF5A mutations , 2014, Neurology.

[24]  Jean-Pierre Julien,et al.  Axonal transport deficits and neurodegenerative diseases , 2013, Nature Reviews Neuroscience.

[25]  Tobias M. Rasse,et al.  Spastic Paraplegia Mutation N256S in the Neuronal Microtubule Motor KIF5A Disrupts Axonal Transport in a Drosophila HSP Model , 2012, PLoS genetics.

[26]  F. Mackenzie,et al.  Crystal Structures of the Tetratricopeptide Repeat Domains of Kinesin Light Chains: Insight into Cargo Recognition Mechanisms , 2012, PloS one.

[27]  A. Ludolph,et al.  Amyotrophic lateral sclerosis. , 2012, Current opinion in neurology.

[28]  Michal Linial,et al.  The complete peptide dictionary – A meta‐proteomics resource , 2010, Proteomics.

[29]  Yi Zhang,et al.  multiplierz: an extensible API based desktop environment for proteomics data analysis , 2009, BMC Bioinformatics.

[30]  J. Milbrandt,et al.  Nicotinamide Mononucleotide Adenylyl Transferase-Mediated Axonal Protection Requires Enzymatic Activity But Not Increased Levels of Neuronal Nicotinamide Adenine Dinucleotide , 2009, The Journal of Neuroscience.

[31]  Jignesh R. Parikh,et al.  mzAPI: a new strategy for efficiently sharing mass spectrometry data , 2009, Nature Methods.

[32]  S. Ficarro,et al.  Improved electrospray ionization efficiency compensates for diminished chromatographic resolution and enables proteomics analysis of tyrosine signaling in embryonic stem cells. , 2009, Analytical chemistry.

[33]  Rebecca Schüle,et al.  Effect of spastic paraplegia mutations in KIF5A kinesin on transport activity. , 2008, Human molecular genetics.

[34]  M. Gorenstein,et al.  Absolute Quantification of Proteins by LCMSE , 2006, Molecular & Cellular Proteomics.

[35]  Nobutaka Hirokawa,et al.  Kinesin Transports RNA Isolation and Characterization of an RNA-Transporting Granule , 2004, Neuron.

[36]  J. Rothnagel,et al.  Alternatively Spliced Products of the Human Kinesin Light Chain 1 (KNS2) Gene , 2003, Traffic.

[37]  David Baltimore,et al.  Germline Transmission and Tissue-Specific Expression of Transgenes Delivered by Lentiviral Vectors , 2002, Science.

[38]  T. Deerinck,et al.  Translocation of RNA Granules in Living Neurons , 1996, The Journal of Neuroscience.

[39]  M A Bisby,et al.  Axonal transport. , 1976, General pharmacology.