A nuclear function of Hu proteins as neuron-specific alternative RNA processing regulators.

Recent advances in genome-wide analysis of alternative splicing indicate that extensive alternative RNA processing is associated with many proteins that play important roles in the nervous system. Although differential splicing and polyadenylation make significant contributions to the complexity of the nervous system, our understanding of the regulatory mechanisms underlying the neuron-specific pathways is very limited. Mammalian neuron-specific embryonic lethal abnormal visual-like Hu proteins (HuB, HuC, and HuD) are a family of RNA-binding proteins implicated in neuronal differentiation and maintenance. It has been established that Hu proteins increase expression of proteins associated with neuronal function by up-regulating mRNA stability and/or translation in the cytoplasm. We report here a novel function of these proteins as RNA processing regulators in the nucleus. We further elucidate the underlying mechanism of this regulation. We show that in neuron-like cells, Hu proteins block the activity of TIA-1/TIAR, two previously identified, ubiquitously expressed proteins that promote the nonneuronal pathway of calcitonin/calcitonin gene-related peptide (CGRP) pre-mRNA processing. These studies define not only the first neuron-specific regulator of the calcitonin/CGRP system but also the first nuclear function of Hu proteins.

[1]  Olivier Pourquié,et al.  Retinoic acid , 2008, Current Biology.

[2]  R. Burry,et al.  HuD Distribution Changes in Response to Heat Shock but Not Neurotrophic Stimulation , 2006, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[3]  Peter Stoilov,et al.  Homologues of the Caenorhabditis elegans Fox-1 Protein Are Neuronal Splicing Regulators in Mammals , 2005, Molecular and Cellular Biology.

[4]  R. Breathnach,et al.  Competition of PTB with TIA proteins for binding to a U-rich cis-element determines tissue-specific splicing of the myosin phosphatase targeting subunit 1. , 2005, RNA.

[5]  K. White,et al.  ELAV Multimerizes on Conserved AU4-6 Motifs Important for ewg Splicing Regulation , 2005, Molecular and Cellular Biology.

[6]  Douglas L. Black,et al.  Polypyrimidine tract binding protein blocks the 5' splice site-dependent assembly of U2AF and the prespliceosomal E complex. , 2005, Molecular cell.

[7]  Robert Castelo,et al.  Regulation of Fas alternative splicing by antagonistic effects of TIA-1 and PTB on exon definition. , 2005, Molecular cell.

[8]  Tyson A. Clark,et al.  Nova regulates brain-specific splicing to shape the synapse , 2005, Nature Genetics.

[9]  D. Lipscombe.  Neuronal proteins custom designed by alternative splicing , 2005, Current Opinion in Neurobiology.

[10]  H. Okano,et al.  The RNA-binding protein HuD regulates neuronal cell identity and maturation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. R. Roesser.  Both U2 snRNA and U12 snRNA are required for accurate splicing of exon 5 of the rat calcitonin/CGRP gene. , 2004, RNA.

[12]  E. Buratti,et al.  An Intronic Polypyrimidine-rich Element Downstream of the Donor Site Modulates Cystic Fibrosis Transmembrane Conductance Regulator Exon 9 Alternative Splicing* , 2004, Journal of Biological Chemistry.

[13]  J. Castle,et al.  Genome-Wide Survey of Human Alternative Pre-mRNA Splicing with Exon Junction Microarrays , 2003, Science.

[14]  Jernej Ule,et al.  CLIP Identifies Nova-Regulated RNA Networks in the Brain , 2003, Science.

[15]  K. White,et al.  ELAV inhibits 3'-end processing to promote neural splicing of ewg pre-mRNA. , 2003, Genes & development.

[16]  H. Lou,et al.  U1 snRNP-Dependent Function of TIAR in the Regulation of Alternative RNA Processing of the Human Calcitonin/CGRP Pre-mRNA , 2003, Molecular and Cellular Biology.

[17]  R. Darnell,et al.  Nova Regulates GABAA Receptor γ2 Alternative Splicing via a Distal Downstream UCAU-Rich Intronic Splicing Enhancer , 2003, Molecular and Cellular Biology.

[18]  J. R. Roesser,et al.  Human transformer 2beta and SRp55 interact with a calcitonin-specific splice enhancer. , 2003, Biochimica et biophysica acta.

[19]  J. R. Roesser,et al.  SRp55 is a regulator of calcitonin/CGRP alternative RNA splicing. , 2003, Biochemistry.

[20]  J. Steitz,et al.  Protein ligands mediate the CRM1-dependent export of HuR in response to heat shock. , 2001, RNA.

[21]  D L Black,et al.  Alternative pre-mRNA splicing and neuronal function. , 2003, Progress in molecular and subcellular biology.

[22]  J. Valcárcel,et al.  The splicing regulator TIA‐1 interacts with U1‐C to promote U1 snRNP recruitment to 5′ splice sites , 2002, The EMBO journal.

[23]  L. Hengst,et al.  ELAV/Hu proteins inhibit p27 translation via an IRES element in the p27 5'UTR. , 2002, Genes & development.

[24]  E. Sakashita,et al.  Complex formation of the neuron-specific ELAV-like Hu RNA-binding proteins. , 2002, Nucleic acids research.

[25]  Christopher J. Lee,et al.  Genome-wide detection of tissue-specific alternative splicing in the human transcriptome. , 2002, Nucleic acids research.

[26]  P. Grabowski,et al.  Region-specific alternative splicing in the nervous system: implications for regulation by the RNA-binding protein NAPOR. , 2002, RNA.

[27]  N. Perrone-Bizzozero,et al.  Role of HuD and other RNA‐binding proteins in neural development and plasticity , 2002, Journal of neuroscience research.

[28]  P. Grabowski,et al.  Function of quaking in myelination: Regulation of alternative splicing , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Christopher J. Lee,et al.  A genomic view of alternative splicing , 2002, Nature Genetics.

[30]  J. Stévenin,et al.  TIA-1 and TIAR Activate Splicing of Alternative Exons with Weak 5′ Splice Sites followed by a U-rich Stretch on Their Own Pre-mRNAs* , 2001, The Journal of Biological Chemistry.

[31]  D. Black,et al.  Alternative RNA splicing in the nervous system , 2001, Progress in Neurobiology.

[32]  J. Qiu,et al.  The neuron-specific RNA-binding protein ELAV regulates neuroglian alternative splicing in neurons and binds directly to its pre-mRNA. , 2001, Genes & development.

[33]  Jack D. Keene,et al.  Ribonucleoprotein infrastructure regulating the flow of genetic information between the genome and the proteome , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[34]  J. A. Steitz,et al.  HuR and mRNA stability , 2001, Cellular and Molecular Life Sciences CMLS.

[35]  J. Valcárcel,et al.  The apoptosis-promoting factor TIA-1 is a regulator of alternative pre-mRNA splicing. , 2000, Molecular cell.

[36]  D. Black,et al.  Cooperative Assembly of an hnRNP Complex Induced by a Tissue-Specific Homolog of Polypyrimidine Tract Binding Protein , 2000, Molecular and Cellular Biology.

[37]  N. Perrone-Bizzozero,et al.  Overexpression of HuD, but Not of Its Truncated Form HuD I+II, Promotes GAP‐43 Gene Expression and Neurite Outgrowth in PC12 Cells in the Absence of Nerve Growth Factor , 2000, Journal of neurochemistry.

[38]  J. Stévenin,et al.  The RNA-Binding Protein TIA-1 Is a Novel Mammalian Splicing Regulator Acting through Intron Sequences Adjacent to a 5′ Splice Site , 2000, Molecular and Cellular Biology.

[39]  R. Darnell,et al.  A brain-enriched polypyrimidine tract-binding protein antagonizes the ability of Nova to regulate neuron-specific alternative splicing. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Robert B Darnell,et al.  Nova-1 Regulates Neuron-Specific Alternative Splicing and Is Essential for Neuronal Viability , 2000, Neuron.

[41]  E. Sakashita,et al.  Cytoplasmic localization is required for the mammalian ELAV‐like protein HuD to induce neuronal differentiation , 1999, Genes to cells : devoted to molecular & cellular mechanisms.

[42]  R. Darnell,et al.  Mammalian ELAV-like neuronal RNA-binding proteins HuB and HuC promote neuronal development in both the central and the peripheral nervous systems. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[43]  J. Keene,et al.  ELAV tumor antigen, Hel-N1, increases translation of neurofilament M mRNA and induces formation of neurites in human teratocarcinoma cells. , 1999, Genes & development.

[44]  H. Lou,et al.  Mechanism of tissue-specific alternative RNA processing of the calcitonin CGRP gene. , 1999, Frontiers of hormone research.

[45]  P. Grabowski,et al.  Coordinate repression of a trio of neuron-specific splicing events by the splicing regulator PTB. , 1999, RNA.

[46]  J. Steitz,et al.  HNS, a nuclear-cytoplasmic shuttling sequence in HuR. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[47]  H. Lou,et al.  Regulation of Alternative Polyadenylation by U1 snRNPs and SRp20 , 1998, Molecular and Cellular Biology.

[48]  M. Ashiya,et al.  A neuron-specific splicing switch mediated by an array of pre-mRNA repressor sites: evidence of a regulatory role for the polypyrimidine tract binding protein and a brain-specific PTB counterpart. , 1997, RNA.

[49]  Y. Wakamatsu,et al.  Sequential expression and role of Hu RNA-binding proteins during neurogenesis. , 1997, Development.

[50]  R. Darnell,et al.  A Hierarchy of Hu RNA Binding Proteins in Developing and Adult Neurons , 1997, The Journal of Neuroscience.

[51]  J. Steitz,et al.  Identification of HuR as a protein implicated in AUUUA‐mediated mRNA decay , 1997, The EMBO journal.

[52]  K. McGowan,et al.  Ectopic expression of Hel-N1, an RNA-binding protein, increases glucose transporter (GLUT1) expression in 3T3-L1 adipocytes , 1997, Molecular and cellular biology.

[53]  S. Cheng,et al.  Purification and Properties of HuD, a Neuronal RNA-binding Protein (*) , 1996, The Journal of Biological Chemistry.

[54]  J. Keene,et al.  Hel-N1/Hel-N2 proteins are bound to poly(A)+ mRNA in granular RNP structures and are implicated in neuronal differentiation. , 1996, Journal of cell science.

[55]  RJ Buckanovich,et al.  The onconeural antigen Nova-1 is a neuron-specific RNA-binding protein, the activity of which is inhibited by paraneoplastic antibodies , 1996, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[56]  H. Lou,et al.  An intron enhancer recognized by splicing factors activates polyadenylation. , 1996, Genes & development.

[57]  H. Lou,et al.  An intron enhancer containing a 5' splice site sequence in the human calcitonin/calcitonin gene-related peptide gene , 1995, Molecular and cellular biology.

[58]  J. Keene,et al.  Hel-N1: an autoimmune RNA-binding protein with specificity for 3' uridylate-rich untranslated regions of growth factor mRNAs. , 1993, Molecular and cellular biology.

[59]  S. Leff,et al.  Regulation of tissue-specific splicing of the calcitonin/calcitonin gene-related peptide gene by RNA-binding proteins. , 1993, Journal of Biological Chemistry.

[60]  M. Malim,et al.  The VP16 transcription activation domain is functional when targeted to a promoter-proximal RNA sequence. , 1992, Genes & development.

[61]  S. Amara,et al.  Molecular Mechanisms of Cell‐Specific and Regulated Expression of the Calcitonin/α‐CGRP and β‐CGRP Genes , 1992 .

[62]  R. Emeson,et al.  Calcitonin Gene—Related Peptide: A Neuropeptide Generated as a Consequence of Tissue‐Specific, Developmentally Regulated Alternative RNA Processing Events , 1992, Annals of the New York Academy of Sciences.

[63]  A. Russo,et al.  Neuronal properties of a thyroid C-cell line: partial repression by dexamethasone and retinoic acid. , 1992, Molecular endocrinology.

[64]  S. Amara,et al.  Molecular mechanisms of cell-specific and regulated expression of the calcitonin/alpha-CGRP and beta-CGRP genes. , 1992, Annals of the New York Academy of Sciences.

[65]  J. Henson,et al.  HuD, a paraneoplastic encephalomyelitis antigen, contains RNA-binding domains and is homologous to Elav and sex-lethal , 1991, Cell.

[66]  G. Adema,et al.  Uridine branch acceptor is a cis-acting element involved in regulation of the alternative processing of calcitonin/CGRP-l pre-mRNA. , 1990, Nucleic acids research.

[67]  S. Leff,et al.  Splice commitment dictates neuron-specific alternative RNA processing in calcitonin/CGRP gene expression , 1987, Cell.

[68]  C. Lips,et al.  Model for alternative RNA processing in human calcitonin gene expression. , 1986, Nucleic acids research.

[69]  J. Davies,et al.  Molecular Biology of the Cell , 1983, Bristol Medico-Chirurgical Journal.

[70]  S. Amara,et al.  Calcitonin mRNA polymorphism: peptide switching associated with alternative RNA splicing events. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[71]  S. Strickland,et al.  The induction of differentiation in teratocarcinoma stem cells by retinoic acid , 1978, Cell.

[72]  H. Studer.  [Paraneoplastic syndromes]. , 1973, Schweizerische medizinische Wochenschrift.