Neurotrophin-Induced Transport of a β-Actin mRNP Complex Increases β-Actin Levels and Stimulates Growth Cone Motility

Abstract Neurotrophin regulation of actin-dependent changes in growth cone motility may depend on the signaling of β-actin mRNA transport. Formation of an RNP complex between the β-actin mRNA zipcode sequence and Zipcode Binding Protein 1 (ZBP1) was required for its localization to growth cones. Antisense oligonucleotides to the zipcode inhibited formation of this RNP complex in vitro and the neurotrophin-induced localization of β-actin mRNA and ZBP1 granules. Live cell imaging of neurons transfected with EGFP-ZBP1 revealed fast, bidirectional movements of granules in neurites that were inhibited by antisense treatment, as visualized by FRAP analysis. NT-3 stimulation of β-actin protein localization was dependent on the 3′UTR and inhibited by antisense treatment. Growth cones exhibited impaired motility in the presense of antisense. These results suggest a novel mechanism to influence growth cone dynamics involving the regulated transport of mRNA.

[1]  A. Hannan,et al.  Intracellular Localization of Tropomyosin mRNA and Protein Is Associated with Development of Neuronal Polarity , 1995, Molecular and Cellular Neuroscience.

[2]  D. Melton,et al.  A two-step model for the localization of maternal mRNA in Xenopus oocytes: involvement of microtubules and microfilaments in the translocation and anchoring of Vg1 mRNA. , 1990, Development.

[3]  Ilan Davis,et al.  Drosophila wingless and Pair-Rule Transcripts Localize Apically by Dynein-Mediated Transport of RNA Particles , 2001, Cell.

[4]  G. Kidd,et al.  Mutational Analysis of a Heterogeneous Nuclear Ribonucleoprotein A2 Response Element for RNA Trafficking* , 1999, The Journal of Biological Chemistry.

[5]  J. Carson,et al.  Translocation of myelin basic protein mRNA in oligodendrocytes requires microtubules and kinesin , 1997 .

[6]  N. Hirokawa,et al.  Kinesin and dynein superfamily proteins and the mechanism of organelle transport. , 1998, Science.

[7]  Xuan Peng,et al.  A Novel Cytoplasmic Protein with RNA-binding Motifs Is an Autoantigen in Human Hepatocellular Carcinoma , 1999, The Journal of experimental medicine.

[8]  K. Shen,et al.  Dynamic control of CaMKII translocation and localization in hippocampal neurons by NMDA receptor stimulation. , 1999, Science.

[9]  R. Singer,et al.  Sequences responsible for intracellular localization of beta-actin messenger RNA also affect cell phenotype , 1994, The Journal of cell biology.

[10]  L. Behar,et al.  Exogenous tau RNA is localized in oocytes: possible evidence for evolutionary conservation of localization mechanisms. , 1996, Developmental biology.

[11]  K. Kosik,et al.  Association of poly(A) mRNA with microtubules in cultured neurons , 1994, Neuron.

[12]  E. Sadot,et al.  cis-Acting signals and trans-acting proteins are involved in tau mRNA targeting into neurites of differentiating neuronal cells , 1995, International Journal of Developmental Neuroscience.

[13]  J. Barg,et al.  Subcellular localization of tau mRNA in differentiating neuronal cell culture: Implications for neuronal polarity , 1993, Neuron.

[14]  O. Steward,et al.  Development of subcellular mRNA compartmentation in hippocampal neurons in culture , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[15]  O. Steward,et al.  Protein synthesis at synaptic sites on dendrites. , 2001, Annual review of neuroscience.

[16]  D. Richter,et al.  Two trans-acting rat-brain proteins, MARTA1 and MARTA2, interact specifically with the dendritic targeting element in MAP2 mRNAs. , 2000, Brain research. Molecular brain research.

[17]  J. Carson,et al.  The Cis-Acting RNA Trafficking Signal from Myelin Basic Protein mRNA and Its Cognate Trans-Acting Ligand Hnrnp A2 Enhance CaP-Dependent Translation , 1999, The Journal of cell biology.

[18]  K. Nasmyth,et al.  Mating type switching in yeast controlled by asymmetric localization of ASH1 mRNA. , 1997, Science.

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

[20]  I. Herman,et al.  Indirect association of ezrin with F-actin: isoform specificity and calcium sensitivity , 1995, The Journal of cell biology.

[21]  R. Singer,et al.  β-Actin Messenger RNA Localization and Protein Synthesis Augment Cell Motility , 1997, The Journal of cell biology.

[22]  K. Kosik,et al.  Neurotrophin-3 signals redistribute RNA in neurons. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[23]  D. Richter,et al.  Identification of a cis-Acting Dendritic Targeting Element in MAP2 mRNAs , 1999, The Journal of Neuroscience.

[24]  N. Hirokawa,et al.  Actin dynamics in growth cones , 1991, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[25]  P. Crino,et al.  Molecular Characterization of the Dendritic Growth Cone: Regulated mRNA Transport and Local Protein Synthesis , 1996, Neuron.

[26]  F. Bloom,et al.  RNA, Whither Goest Thou? , 1999, Science.

[27]  J. Barg,et al.  Microtubules are involved in the localization of tau mRNA in primary neuronal cell cultures , 1994, Neuron.

[28]  J. Brosius,et al.  Translational Machinery in Dendrites of Hippocampal Neurons in Culture , 1996, Journal of Neuroscience.

[29]  C. Nüsslein-Volhard,et al.  The molecular motor dynein is involved in targeting Swallow and bicoid RNA to the anterior pole of Drosophila oocytes , 2000, Nature Cell Biology.

[30]  K. Kosik,et al.  Sorting of β-Actin mRNA and Protein to Neurites and Growth Cones in Culture , 1998, The Journal of Neuroscience.

[31]  J. Condeelis,et al.  The physiological significance of β-actin mRNA localization in determining cell polarity and directional motility , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[32]  R. Singer,et al.  Neuronal RNA localization and the cytoskeleton. , 2001, Results and problems in cell differentiation.

[33]  R. Singer,et al.  A Rho-dependent signaling pathway operating through myosin localizes β-actin mRNA in fibroblasts , 2001, Current Biology.

[34]  R. Singer,et al.  Requirement of microfilaments in sorting of actin messenger RNA. , 1991, Science.

[35]  G. Gallo,et al.  Localized Sources of Neurotrophins Initiate Axon Collateral Sprouting , 1998, The Journal of Neuroscience.

[36]  M. Kiebler,et al.  Microtubule-dependent recruitment of Staufen-green fluorescent protein into large RNA-containing granules and subsequent dendritic transport in living hippocampal neurons. , 1999, Molecular biology of the cell.

[37]  J. Bamburg,et al.  Regulating actin dynamics in neuronal growth cones by ADF/cofilin and rho family GTPases. , 2000, Journal of neurobiology.

[38]  K. Kosik,et al.  CaMKIIα 3′ Untranslated Region-Directed mRNA Translocation in Living Neurons: Visualization by GFP Linkage , 2000, The Journal of Neuroscience.

[39]  A. Git,et al.  RNA-binding protein conserved in both microtubule- and microfilament-based RNA localization. , 1998, Genes & development.

[40]  M. Kiebler,et al.  The Mammalian Staufen Protein Localizes to the Somatodendritic Domain of Cultured Hippocampal Neurons: Implications for Its Involvement in mRNA Transport , 1999, The Journal of Neuroscience.

[41]  E. Kandel,et al.  The 3'-untranslated region of CaMKII alpha is a cis-acting signal for the localization and translation of mRNA in dendrites. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[42]  K. Hoek,et al.  hnRNP A2 selectively binds the cytoplasmic transport sequence of myelin basic protein mRNA. , 1998, Biochemistry.

[43]  S. Kaech,et al.  Improved lipid-mediated gene transfer into primary cultures of hippocampal neurons. , 1996, Brain research. Molecular brain research.

[44]  Daniela Avossa,et al.  Transport and Localization Elements in Myelin Basic Protein mRNA , 1997, The Journal of cell biology.

[45]  I. Ringel,et al.  Vg1 RNA binding protein mediates the association of Vg1 RNA with microtubules in Xenopus oocytes. , 1995, The EMBO journal.

[46]  R. Singer,et al.  Neurotrophin Regulation of β-Actin mRNA and Protein Localization within Growth Cones , 1999, The Journal of cell biology.

[47]  R. Singer,et al.  Localization of ASH1 mRNA particles in living yeast. , 1998, Molecular cell.

[48]  J. Carson,et al.  Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes , 1993, The Journal of cell biology.

[49]  W. Saxton,et al.  A function for kinesin I in the posterior transport of oskar mRNA and Staufen protein. , 2000, Science.

[50]  M. Kiebler,et al.  Molecular Insights into mRNA Transport and Local Translation in the Mammalian Nervous System , 2000, Neuron.

[51]  James O. Deshler,et al.  A highly conserved RNA-binding protein for cytoplasmic mRNA localization in vertebrates , 1998, Current Biology.

[52]  M. Saarma,et al.  Neurotrophins as in vitro growth cone guidance molecules for embryonic sensory neurons , 1997, Cell and Tissue Research.

[53]  M. Tohyama,et al.  Two cis-acting elements in the 3′ untranslated region of α-CaMKII regulate its dendritic targeting , 2000, Nature Neuroscience.

[54]  R. Singer,et al.  Characterization of a beta-actin mRNA zipcode-binding protein , 1997, Molecular and cellular biology.