DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: NELL2-Robo3 complex structure reveals mechanisms of receptor activation for axon guidance

Journal Article · · Nature Communications

Axon pathfinding is critical for nervous system development, and it is orchestrated by molecular cues that activate receptors on the axonal growth cone. Robo family receptors bind Slit guidance cues to mediate axon repulsion. In mammals, the divergent family member Robo3 does not bind Slits, but instead signals axon repulsion from its own ligand, NELL2. Conversely, canonical Robos do not mediate NELL2 signaling. Here, we present the structures of NELL-Robo3 complexes, identifying a mode of ligand engagement for Robos that is orthogonal to Slit binding. We elucidate the structural basis for differential binding between NELL and Robo family members and show that NELL2 repulsive activity is a function of its Robo3 affinity and is enhanced by ligand trimerization. Our results reveal a mechanism of oligomerization-induced Robo activation for axon guidance and shed light on Robo family member ligand binding specificity, conformational variability, divergent modes of signaling, and evolution.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS); USDOE Office of Science (SC)
Grant/Contract Number:
AC02-06CH11357; T32 GM077995; R01 NS097161; R01 MH114102; RI-INBRE P20 GM103430; R01 NS095908; S10 RR029205; 1S10OD012289-01A1; 9 P41 GM103622; 1S10OD018090-01
OSTI ID:
1608059
Journal Information:
Nature Communications, Vol. 11, Issue 1; ISSN 2041-1723
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 21 works
Citation information provided by
Web of Science

References (91)

Uniqueness of ab initio shape determination in small-angle scattering journal April 2003
Structural Divergence in O-GlcNAc Glycans Displayed on Epidermal Growth Factor-like Repeats of Mammalian Notch1 journal July 2018
DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering journal January 2009
Slit-Robo signaling journal August 2016
Regulation of Commissural Axon Pathfinding by Slit and its Robo Receptors journal November 2006
BioXTAS RAW : improvements to a free open-source program for small-angle X-ray scattering data reduction and analysis journal September 2017
Structural insights into the Slit-Robo complex journal September 2007
TheDUTT1Gene, a Novel NCAM Family Member Is Expressed in Developing Murine Neural Tissues and Has an Unusually Broad Pattern of Expression journal May 1998
New gene, nel, encoding a Mr 93 K protein with EGF-like repeats is strongly expressed in neural tissues of early stage chick embryos journal June 1995
Nell2 regulates the contralateral-versus-ipsilateral visual projection as a domain-specific positional cue journal February 2019
Signaling Switch of the Axon Guidance Receptor Robo3 during Vertebrate Evolution journal December 2014
Netrins are diffusible chemotropic factors for commissural axons in the embryonic spinal cord journal August 1994
Sonic Hedgehog Guides Axons through a Noncanonical, Src-Family-Kinase-Dependent Signaling Pathway journal May 2009
Towards automated crystallographic structure refinement with phenix.refine journal March 2012
Extracellular Ig domains 1 and 2 of Robo are important for ligand (Slit) binding journal June 2004
Overview of Current Methods in Sedimentation Velocity and Sedimentation Equilibrium Analytical Ultracentrifugation journal February 2013
Robo Ig4 Is a Dimerization Domain journal November 2017
Alternative Splicing of the Robo3 Axon Guidance Receptor Governs the Midline Switch from Attraction to Repulsion journal May 2008
Magic Roundabout Is a New Member of the Roundabout Receptor Family That Is Endothelial Specific and Expressed at Sites of Active Angiogenesis journal April 2002
Robo family of proteins exhibit differential expression in mouse spinal cord and Robo-Slit interaction is required for midline crossing in vertebrate spinal cord journal January 2005
Decision-making in structure solution using Bayesian estimates of map quality: the PHENIX AutoSol wizard journal May 2009
Structure and functional relevance of the Slit2 homodimerization domain journal May 2009
Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells journal September 2007
Biochemical Characterization and Expression Analysis of Neural Thrombospondin-1-like Proteins NELL1 and NELL2 journal November 1999
Restricted neural epidermal growth factor-like like 2 (NELL2) expression during muscle and neuronal differentiation journal December 2002
Structures of netrin-1 bound to two receptors provide insight into its axon guidance mechanism journal May 2014
Size-Distribution Analysis of Macromolecules by Sedimentation Velocity Ultracentrifugation and Lamm Equation Modeling journal March 2000
Revisiting Netrin-1: One Who Guides (Axons) journal July 2018
Structural Principles in Robo Activation and Auto-inhibition journal April 2019
XDS journal January 2010
Mutations in a Human ROBO Gene Disrupt Hindbrain Axon Pathway Crossing and Morphogenesis journal June 2004
Tandem mass spectrometry identifies many mouse brain O-GlcNAcylated proteins including EGF domain-specific O-GlcNAc transferase targets journal April 2012
MoRDa , an automatic molecular replacement pipeline journal August 2015
The Crystal Structure of Netrin-1 in Complex with DCC Reveals the Bifunctionality of Netrin-1 As a Guidance Cue journal August 2014
Robo1 Forms a Compact Dimer-of-Dimers Assembly journal February 2018
Nell1-deficient mice have reduced expression of extracellular matrix proteins causing cranial and vertebral defects journal March 2006
Ontogeny and the possible function of a novel epidermal growth factor-like repeat domain-containing protein, NELL2, in the rat brain: A role of NELL2 in the rat brain journal December 2002
Mechanisms and Molecules of Neuronal Wiring: A Primer journal December 2010
Cloning and Characterization of Two Novel Human cDNAs (NELL1 and NELL2) Encoding Proteins with Six EGF-like Repeats journal December 1996
MolProbity : all-atom structure validation for macromolecular crystallography journal December 2009
Evidence for the existence of two Robo3 isoforms with divergent biochemical properties journal December 2005
Features and development of Coot journal March 2010
Cell-surface heparan sulfate is involved in the repulsive guidance activities of Slit2 protein journal July 2001
Calculations and Publication-Quality Illustrations for Analytical Ultracentrifugation Data book January 2015
Molecular Logic of Neuronal Self-Recognition through Protocadherin Domain Interactions journal October 2015
Structural Basis for Regulation of GPR56/ADGRG1 by Its Alternatively Spliced Extracellular Domains journal September 2016
In vitro guidance of retinal axons by a tectal lamina-specific glycoprotein Nel journal June 2009
An Extracellular Interactome of Immunoglobulin and LRR Proteins Reveals Receptor-Ligand Networks journal July 2013
Binding site for Robo receptors revealed by dissection of the leucine-rich repeat region of Slit journal October 2004
Accurate assessment of mass, models and resolution by small-angle scattering journal April 2013
slit: an extracellular protein necessary for development of midline glia and commissural axon pathways contains both EGF and LRR domains. journal December 1990
Operational redundancy in axon guidance through the multifunctional receptor Robo3 and its ligand NELL2 journal November 2015
Phaser crystallographic software journal July 2007
Protocadherin cis -dimer architecture and recognition unit diversity journal October 2017
New gene, nel, encoding a Mr 93 K protein with EGF-like repeats is strongly expressed in neural tissues of early stage chick embryos journal June 1995
Robo family of proteins exhibit differential expression in mouse spinal cord and Robo-Slit interaction is required for midline crossing in vertebrate spinal cord journal January 2005
Biochemical Characterization and Expression Analysis of Neural Thrombospondin-1-like Proteins NELL1 and NELL2 journal November 1999
Cloning and Characterization of Two Novel Human cDNAs (NELL1 and NELL2) Encoding Proteins with Six EGF-like Repeats journal December 1996
Magic Roundabout Is a New Member of the Roundabout Receptor Family That Is Endothelial Specific and Expressed at Sites of Active Angiogenesis journal April 2002
Molecular Logic of Neuronal Self-Recognition through Protocadherin Domain Interactions journal October 2015
Structural Principles in Robo Activation and Auto-inhibition journal April 2019
Robo Ig4 Is a Dimerization Domain journal November 2017
In vitro guidance of retinal axons by a tectal lamina-specific glycoprotein Nel journal June 2009
To Cross or Not to Cross: Alternatively Spliced Forms of the Robo3 Receptor Regulate Discrete Steps in Axonal Midline Crossing journal May 2008
The Crystal Structure of Netrin-1 in Complex with DCC Reveals the Bifunctionality of Netrin-1 As a Guidance Cue journal August 2014
Signaling Switch of the Axon Guidance Receptor Robo3 during Vertebrate Evolution journal December 2014
Structural Basis for Regulation of GPR56/ADGRG1 by Its Alternatively Spliced Extracellular Domains journal September 2016
Slit Proteins Bind Robo Receptors and Have an Evolutionarily Conserved Role in Repulsive Axon Guidance journal March 1999
Conservation and divergence of axon guidance mechanisms journal October 1999
Cell-surface heparan sulfate is involved in the repulsive guidance activities of Slit2 protein journal July 2001
Accurate assessment of mass, models and resolution by small-angle scattering journal April 2013
Binding site for Robo receptors revealed by dissection of the leucine-rich repeat region of Slit journal October 2004
Roundabout 1 exists predominantly as a basal dimeric complex and this is unaffected by binding of the ligand Slit2 journal June 2014
Structural insight into Slit–Robo signalling journal March 2008
Ontogeny and the possible function of a novel epidermal growth factor-like repeat domain-containing protein, NELL2, in the rat brain: A role of NELL2 in the rat brain journal December 2002
Structural insights into the Slit-Robo complex journal September 2007
Oligomerization-induced Conformational Change in the C-terminal Region of Nel-like Molecule 1 (NELL1) Protein Is Necessary for the Efficient Mediation of Murine MC3T3-E1 Cell Adhesion and Spreading journal February 2014
A Molecular Mechanism for the Heparan Sulfate Dependence of Slit-Robo Signaling journal October 2006
Structural and Functional Analysis of Slit and Heparin Binding to Immunoglobulin-like Domains 1 and 2 of Drosophila Robo journal March 2008
Robo2 contains a cryptic binding site for neural EGFL-like (NELL) protein 1/2 journal January 2019
Nell1-deficient mice have reduced expression of extracellular matrix proteins causing cranial and vertebral defects journal March 2006
Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells journal September 2007
Structures of netrin-1 bound to two receptors provide insight into its axon guidance mechanism journal May 2014
Regulation of Commissural Axon Pathfinding by Slit and its Robo Receptors journal November 2006
Role of ROBO4 Signalling in Developmental and Pathological Angiogenesis journal January 2014
Slit-Robo signaling journal August 2016
Nell2 regulates the contralateral-versus-ipsilateral visual projection as a domain-specific positional cue journal February 2019
Collaborative and Specialized Functions of Robo1 and Robo2 in Spinal Commissural Axon Guidance journal July 2010
Decision-making in structure solution using Bayesian estimates of map quality: the PHENIX AutoSol wizard. text January 2009
Robo2 acts in trans to inhibit Slit-Robo1 repulsion in pre-crossing commissural axons journal July 2015
Molecular basis of synaptic specificity by immunoglobulin superfamily receptors in Drosophila journal January 2019

Cited By (91)

Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells journal August 2019
A General Approach to Access Morphologies of Polyoxometalates in Solution by Using SAXS: An Ab Initio Modeling Protocol journal April 2018
Intrinsic structural disorder of mouse proNGF journal June 2009
Biochemical characterization and low-resolution SAXS structure of two-domain endoglucanase BlCel9 from Bacillus licheniformis journal December 2018
WeNMR: Structural Biology on the Grid journal November 2012
Structural Rearrangements in the C-Terminal Domain Homolog of Orange Carotenoid Protein are Crucial for Carotenoid Transfer journal February 2019
Structural peculiarities of the (MHF1-MHF2) 4 octamer provide a long DNA binding patch to anchor the MHF-FANCM complex to chromatin: A solution SAXS study journal July 2013
Structure of a Novel Dimeric SET Domain Methyltransferase that Regulates Cell Motility journal October 2018
Structural basis for iron piracy by pathogenic Neisseria journal February 2012
Structural basis for the assembly of the Sxl–Unr translation regulatory complex journal September 2014
Molecular architecture of the human sperm IZUMO1 and egg JUNO fertilization complex journal June 2016
Structural and mechanistic insights into regulation of the retromer coat by TBC1d5 journal November 2016
AAA+ chaperones and acyldepsipeptides activate the ClpP protease via conformational control journal February 2015
Molecular basis of human CD22 function and therapeutic targeting journal October 2017
A viral-fusion-peptide-like molecular switch drives membrane insertion of botulinum neurotoxin A1 journal December 2018
Unique structural features of a bacterial autotransporter adhesin suggest mechanisms for interaction with host macromolecules journal April 2019
Mycobacterium tuberculosis CarD, an essential global transcriptional regulator forms amyloid-like fibrils journal July 2018
Molecular architecture of the Jumonji C family histone demethylase KDM5B journal March 2019
Nano-encapsulated Escherichia coli Divisome Anchor ZipA, and in Complex with FtsZ journal December 2019
ClpP protease activation results from the reorganization of the electrostatic interaction networks at the entrance pores journal November 2019
Structural basis of a nucleosome containing histone H2A.B/H2A.Bbd that transiently associates with reorganized chromatin journal December 2013
Crystal structure of Streptococcus pneumoniae pneumolysin provides key insights into early steps of pore formation journal September 2015
Disease-Homologous Mutation in the Cation Diffusion Facilitator Protein MamM Causes Single-Domain Structural Loss and Signifies Its Importance journal August 2016
The N-terminal domain of unknown function (DUF959) in collagen XVIII is intrinsically disordered and highly O-glycosylated journal November 2018
Conserved structure and domain organization among bacterial Slc26 transporters journal September 2014
Light-activated DNA binding in a designed allosteric protein journal July 2008
The human IgM pentamer is a mushroom-shaped molecule with a flexural bias journal August 2009
Two crystal structures reveal design for repurposing the C-Ala domain of human AlaRS journal November 2016
How members of the human gut microbiota overcome the sulfation problem posed by glycosaminoglycans journal June 2017
DC-SIGN Neck Domain Is a pH-sensor Controlling Oligomerization: SAXS AND HYDRODYNAMIC STUDIES OF EXTRACELLULAR DOMAIN journal June 2009
Electrostatic Interactions Involving the Extreme C Terminus of Nuclear Export Factor CRM1 Modulate Its Affinity for Cargo journal August 2011
Fission Yeast Swi5-Sfr1 Protein Complex, an Activator of Rad51 Recombinase, Forms an Extremely Elongated Dogleg-shaped Structure journal December 2011
Interaction of the HIV-1 Intasome with Transportin 3 Protein (TNPO3 or TRN-SR2) journal October 2012
Characterization of Enzymes from Legionella pneumophila Involved in Reversible Adenylylation of Rab1 Protein journal October 2012
Differential Role of HAMP-like Linkers in Regulating the Functionality of the Group III Histidine Kinase DhNik1p journal July 2014
Assembly of the Respiratory Mucin MUC5B journal June 2014
Structural Basis for Oxygen Activation at a Heterodinuclear Manganese/Iron Cofactor journal October 2015
Solution Conformation and Thermodynamic Characteristics of RNA Binding by the Splicing Factor U2AF 65 journal October 2008
Structure-specific recognition protein-1 (SSRP1) is an elongated homodimer that binds histones journal June 2018
The antibiotic cyclomarin blocks arginine-phosphate–induced millisecond dynamics in the N-terminal domain of ClpC1 from Mycobacterium tuberculosis journal June 2018
Direct observation of conformational dynamics of the PH domain in phospholipases Cɛ and β may contribute to subfamily-specific roles in regulation journal November 2018
The dimeric ectodomain of the alkali-sensing insulin receptor–related receptor (ectoIRR) has a droplike shape journal November 2019
Integrative Structure–Function Mapping of the Nucleoporin Nup133 Suggests a Conserved Mechanism for Membrane Anchoring of the Nuclear Pore Complex journal August 2014
Disease-associated mutations alter the dynamic motion of the N-terminal domain of the human cardiac ryanodine receptor journal March 2019
Simple rules for passive diffusion through the nuclear pore complex journal October 2016
Structural evaluation of an amyloid fibril model using small-angle x-ray scattering journal June 2017
Calcium-dependent structural changes in human reticulocalbin-1 journal January 2014
Quaternary structure of the yeast Arc1p-aminoacyl-tRNA synthetase complex in solution and its compaction upon binding of tRNAs journal November 2012
The H-subunit of the restriction endonuclease CglI contains a prototype DEAD-Z1 helicase-like motor journal February 2018
Structure of human telomere G-quadruplex in the presence of a model drug along the thermal unfolding pathway journal November 2018
Structure and hydrodynamics of a DNA G-quadruplex with a cytosine bulge journal May 2018
Microstructural degeneracy associated with a two-point correlation function and its information content journal May 2012
Combination of X-ray crystallography, SAXS and DEER to obtain the structure of the FnIII-3,4 domains of integrin α6β4 journal March 2015
Interpretation of solution scattering data from lipid nanodiscs journal February 2018
Photocage-initiated time-resolved solution X-ray scattering investigation of protein dimerization journal September 2018
The structure of VgrG1 from Pseudomonas aeruginosa , the needle tip of the bacterial type VI secretion system journal January 2016
Structural insights into the interaction of the conserved mammalian proteins GAPR-1 and Beclin 1, a key autophagy protein journal August 2017
Solution structures of long-acting insulin analogues and their complexes with albumin text January 2019
Flexibility of truncated and full-length glucansucrase GTF180 enzymes from Lactobacillus reuteri 180 journal March 2014
Borrelia outer surface protein C is capable of human fibrinogen binding journal March 2019
A biophysical and structural study of two chitinases from Agave tequilana and their potential role as defense proteins journal July 2019
GGA Autoinhibition Revisited journal February 2010
Antagonists Induce a Conformational Change in cIAP1 That Promotes Autoubiquitination journal October 2011
Structural Basis for Assembly and Function of a Heterodimeric Plant Immune Receptor journal April 2014
Solution Structure, Self-Assembly, and Membrane Interactions of the Matrix Protein from Newcastle Disease Virus at Neutral and Acidic pH journal March 2019
Molecular Basis of Unusually High Neutralization Resistance in Tier 3 HIV-1 Strain 253-11 journal April 2018
Структура частиц белка оболочки А вируса картофеля и их диссоциация journal January 2018
High-resolution structure of a type IV pilin from the metal-reducing bacterium Shewanella oneidensis journal February 2015
The mechanism by which a distinguishing arabinofuranosidase can cope with internal di-substitutions in arabinoxylans journal August 2018
Domain Swapping and Different Oligomeric States for the Complex Between Calmodulin and the Calmodulin-Binding Domain of Calcineurin A journal April 2009
Structural Insights into the Mechanism of Phosphoregulation of the Retinoblastoma Protein journal March 2013
Low Resolution Structural Studies Indicate that the Activator of Hsp90 ATPase 1 (Aha1) of Leishmania braziliensis Has an Elongated Shape Which Allows Its Interaction with Both N- and M-Domains of Hsp90 journal June 2013
Structural Analysis of Influenza A Virus Matrix Protein M1 and Its Self-Assemblies at Low pH journal December 2013
An alkaline active feruloyl-CoA synthetase from soil metagenome as a potential key enzyme for lignin valorization strategies journal February 2019
The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces journal January 2012
Lectin-Like Bacteriocins from Pseudomonas spp. Utilise D-Rhamnose Containing Lipopolysaccharide as a Cellular Receptor journal February 2014
Long non‐coding subgenomic flavivirus RNAs have extended 3D structures and are flexible in solution journal September 2019
Maturation of 6S regulatory RNA to a highly elongated structure text January 2015
Structural and Dynamics Studies of a Truncated Variant of CI Repressor From Bacteriophage TP901-1 text January 2016
The Redox State Regulates the Conformation of Rv2466c to Activate the Antitubercular Prodrug TP053 text January 2015
X-Ray Solution Scattering Study of Four Escherichia coli Enzymes Involved in Stationary-Phase Metabolism text January 2016
VirB8-like protein TraH is crucial for DNA transfer in Enterococcus faecalis text January 2016
Myelin 2′,3′-Cyclic Nucleotide 3′-Phosphodiesterase: Active-Site Ligand Binding and Molecular Conformation text January 2012
Molecular basis of the 14-3-3 protein-dependent activation of yeast neutral trehalase Nth1 text January 2017
SAXS and stability studies of iron-induced oligomers of bacterial frataxin CyaY text January 2017
Conformational characterization of full-length X-chromosome-linked inhibitor of apoptosis protein (XIAP) through an integrated approach text January 2019
Conformational States of Exchange Protein Directly Activated by cAMP (EPAC1) Revealed by Ensemble Modeling and Integrative Structural Biology journal December 2019
Combination of SAXS and Protein Painting Discloses the Three-Dimensional Organization of the Bacterial Cysteine Synthase Complex, a Potential Target for Enhancers of Antibiotic Action journal October 2019
Review of the fundamental theories behind small angle X-ray scattering, molecular dynamics simulations, and relevant integrated application journal January 2015
Phosphorylation-mediated PTEN conformational closure and deactivation revealed with protein semisynthesis journal July 2013
Importin-9 wraps around the H2A-H2B core to act as nuclear importer and histone chaperone journal March 2019

Similar Records

Tau mediates microtubule bundle architectures mimicking fascicles of microtubules found in the axon initial segment
Journal Article · 2016 · Nature Communications · OSTI ID:1326678

Structure and Activity of the Axon Guidance Protein MICAL
Journal Article · 2005 · Proc Natl Acad Sci USA · OSTI ID:913715

Family of neural wiring receptors in bilaterians defined by phylogenetic, biochemical, and structural evidence
Journal Article · 2019 · Proceedings of the National Academy of Sciences of the United States of America · OSTI ID:1526067

Related Subjects