Structural bases for CRMP function in plexin‐dependent semaphorin3A signaling

Collapsin response mediator proteins (CRMPs) are cytosolic phosphoproteins involved in neuronal differentiation and axonal guidance. CRMP2 was previously shown to mediate the repulsive effect of Sema3A on axons and to participate in axonal specification. The X‐ray crystal structure of murine CRMP1 was determined at 2.1 Å resolution and demonstrates that CRMP1 is a bilobed ‘lung‐shaped’ protein forming a tetrameric assembly. Structure‐based mutagenesis of surface‐exposed residues was employed to map functional domains. As a rapid assay for CRMP, we exploited a reconstituted Sema3A signaling system in COS‐7 cells expressing the receptor components Neuropilin1 and PlexinA1 (NP1/PlexA1). In these cells, CRMP and PlexA1 form a physical complex that is reduced in amount by NP1 but enhanced by Sema3A/NP1. Furthermore, CRMP accelerates Sema3A‐induced cell contraction. Alanine substitutions in one domain of CRMP1 produce a constitutively active protein that causes Sema3A‐independent COS‐7 contraction. This mutant CRMP mimics the DRG neurite outgrowth‐inhibiting effects of Sema3A and reduces Sema3A‐induced axonal repulsion. These data provide a structural view of CRMP function in Plex‐dependent Sema3A signaling.

[1]  C. Sander,et al.  Database of homology‐derived protein structures and the structural meaning of sequence alignment , 1991, Proteins.

[2]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[3]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[4]  R. K. Herman,et al.  Analysis of the Caenorhabditis elegans axonal guidance and outgrowth gene unc-33. , 1992, Genetics.

[5]  A. Brünger.  Free R value: a novel statistical quantity for assessing the accuracy of crystal structures , 1992, Nature.

[6]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[7]  V S Lamzin,et al.  Automated refinement of protein models. , 1993, Acta crystallographica. Section D, Biological crystallography.

[8]  C. Sander,et al.  Protein structure comparison by alignment of distance matrices. , 1993, Journal of molecular biology.

[9]  T. Takagi,et al.  Purification, characterization and inhibition of dihydropyrimidinase from rat liver. , 1994, European journal of biochemistry.

[10]  S. Hockfield,et al.  TOAD-64, a gene expressed early in neuronal differentiation in the rat, is related to unc-33, a C. elegans gene involved in axon outgrowth , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[11]  F. Raushel,et al.  Three-dimensional structure of the binuclear metal center of phosphotriesterase. , 1995, Biochemistry.

[12]  Fumio Nakamura,et al.  Collapsin-induced growth cone collapse mediated by an intracellular protein related to UNC-33 , 1995, Nature.

[13]  S. Grzesiek,et al.  NMRPipe: A multidimensional spectral processing system based on UNIX pipes , 1995, Journal of biomolecular NMR.

[14]  S. Ciurli,et al.  X-ray absorption spectroscopy study of native and phenylphosphorodiamidate-inhibited Bacillus pasteurii urease. , 1996, European journal of biochemistry.

[15]  S. Strittmatter,et al.  Rac1 Mediates Collapsin-1-Induced Growth Cone Collapse , 1997, The Journal of Neuroscience.

[16]  M. Tessier-Lavigne,et al.  Neuropilin Is a Receptor for the Axonal Chemorepellent Semaphorin III , 1997, Cell.

[17]  E J Dodson,et al.  Collaborative Computational Project, number 4: providing programs for protein crystallography. , 1997, Methods in enzymology.

[18]  Alex L Kolodkin,et al.  Neuropilin Is a Semaphorin III Receptor , 1997, Cell.

[19]  T A Jones,et al.  Electron-density map interpretation. , 1997, Methods in enzymology.

[20]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[21]  C Sander,et al.  An evolutionary treasure: unification of a broad set of amidohydrolases related to urease , 1997, Proteins.

[22]  S. Strittmatter,et al.  Brain CRMP Forms Heterotetramers Similar to Liver Dihydropyrimidinase , 1997, Journal of neurochemistry.

[23]  V. N. Molchanov,et al.  Superconducting Single Crystals of Tl2Ba2CaCu2O8 and YBa2Cu4O8: Crystal Structures in the Vicinity of Tc , 1998 .

[24]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[25]  F A Quiocho,et al.  Complexes of adenosine deaminase with two potent inhibitors: X-ray structures in four independent molecules at pH of maximum activity. , 1998, Biochemistry.

[26]  J. Honnorat,et al.  Ulip/CRMP proteins are recognized by autoantibodies in paraneoplastic neurological syndromes , 1999, The European journal of neuroscience.

[27]  Russ Miller,et al.  The design and implementation of SnB version 2.0 , 1999 .

[28]  R M Esnouf,et al.  Further additions to MolScript version 1.4, including reading and contouring of electron-density maps. , 1999, Acta crystallographica. Section D, Biological crystallography.

[29]  R. Kalb,et al.  Plexin-Neuropilin-1 Complexes Form Functional Semaphorin-3A Receptors , 1999, Cell.

[30]  M. Noda,et al.  Molecular Characterization of CRMP5, a Novel Member of the Collapsin Response Mediator Protein Family* , 2000, The Journal of Biological Chemistry.

[31]  Jonathan A Raper,et al.  Semaphorins and their receptors in vertebrates and invertebrates , 2000, Current Opinion in Neurobiology.

[32]  R. Kalb,et al.  Molecular basis of semaphorin-mediated axon guidance. , 2000, Journal of neurobiology.

[33]  S. Yanagi,et al.  Identification of CRAM, a novel unc-33 gene family protein that associates with CRMP3 and protein-tyrosine kinase(s) in the developing rat brain. , 2000, The Journal of biological chemistry.

[34]  M. Poo,et al.  Plexins Are a Large Family of Receptors for Transmembrane, Secreted, and GPI-Anchored Semaphorins in Vertebrates , 1999, Cell.

[35]  Y. Goshima,et al.  Phosphorylation of cofilin by LIM-kinase is necessary for semaphorin 3A-induced growth cone collapse , 2001, Nature Neuroscience.

[36]  M. Tessier-Lavigne,et al.  Plexin-A3 Mediates Semaphorin Signaling and Regulates the Development of Hippocampal Axonal Projections , 2001, Neuron.

[37]  H M Holden,et al.  Molecular structure of dihydroorotase: a paradigm for catalysis through the use of a binuclear metal center. , 2001, Biochemistry.

[38]  S. Strittmatter,et al.  PlexinA1 Autoinhibition by the Plexin Sema Domain , 2001, Neuron.

[39]  K. Kaibuchi,et al.  CRMP-2 induces axons in cultured hippocampal neurons , 2001, Nature Neuroscience.

[40]  D. Schomburg,et al.  The structure of L-hydantoinase from Arthobacter aurescens leads to an understanding of dihydropyrimidinase substrate and enantio specificity. , 2002, Biochemistry.

[41]  Y. Goshima,et al.  Collapsin Response Mediator Protein-2 Inhibits Neuronal Phospholipase D2 Activity by Direct Interaction* , 2002, The Journal of Biological Chemistry.

[42]  K. Kaibuchi,et al.  CRMP-2 binds to tubulin heterodimers to promote microtubule assembly , 2002, Nature Cell Biology.

[43]  D. Schomburg,et al.  Crystal structure of D-hydantoinase from Bacillus stearothermophilus: insight into the stereochemistry of enantioselectivity. , 2002, Biochemistry.

[44]  Gerry McDermott,et al.  The structure of Escherichia coli cytosine deaminase. , 2002, Journal of molecular biology.

[45]  Dietmar Schomburg,et al.  X-ray structure of a dihydropyrimidinase from Thermus sp. at 1.3 A resolution. , 2002, Journal of molecular biology.

[46]  Y. Goshima,et al.  Involvement of Fes/Fps tyrosine kinase in semaphorin3A signaling , 2002, The EMBO journal.

[47]  S. Hockfield,et al.  TUC-4b, a Novel TUC Family Variant, Regulates Neurite Outgrowth and Associates with Vesicles in the Growth Cone , 2003, The Journal of Neuroscience.

[48]  Yunqing Liu,et al.  Crystal Structure of d-Hydantoinase from Burkholderia pickettii at a Resolution of 2.7 Angstroms: Insights into the Molecular Basis of Enzyme Thermostability , 2003, Journal of bacteriology.