Disabled-1 interacts with a novel developmentally regulated protocadherin.

Disabled-1 (Dab1) is an intracellular adapter protein that mediates the effect of Reelin on neuronal migration and cell positioning during mammalian brain development. To identify components of the Reelin-Dab1 signaling pathway, we searched for proteins that interact with Dab1 using a yeast two-hybrid strategy. We found that the Dab1 phosphotyrosine binding (PTB) domain interacts with a novel protocadherin, orthologous to human protocadherin 18. Mouse Pcdh18 (mPcdh18), which consists of four exons similar to other protocadherin family members, maps to chromosome 3. The deduced amino acid sequence of mPcdh18 contains six extracellular cadherin motifs, a single transmembrane region, and a large intracellular domain. The site of Dab1 interaction was localized to the C-terminal 243 residues of mPcdh18. Expression analyses revealed that mPcdh18 is present in a variety of tissues in the embryo, but in adult mice it is primarily expressed in lung and kidney. In embryonic brain, mPcdh18 expression is temporally and spatially regulated. Our results indicate that mPcdh18 participates in signaling pathways involving PTB-containing proteins and suggest that it may play a role during brain development.

[1]  Peter D. Kwong,et al.  Structural basis of cell-cell adhesion by cadherins , 1995, Nature.

[2]  T. Curran,et al.  Disabled-1 acts downstream of Reelin in a signaling pathway that controls laminar organization in the mammalian brain. , 1998, Development.

[3]  Amy S. Espeseth,et al.  NF-protocadherin, a novel member of the cadherin superfamily, is required for Xenopus ectodermal differentiation , 1998, Current Biology.

[4]  C. G. Wright,et al.  The mouse Ames waltzer hearing-loss mutant is caused by mutation of Pcdh15, a novel protocadherin gene , 2001, Nature Genetics.

[5]  Q. Al-Awqati,et al.  μ-Protocadherin, a Novel Developmentally Regulated Protocadherin with Mucin-like Domains* , 2000, The Journal of Biological Chemistry.

[6]  T. Pawson,et al.  The PTB domain: a new protein module implicated in signal transduction. , 1995, Trends in biochemical sciences.

[7]  Jonathan A. Cooper,et al.  Dab1 tyrosine phosphorylation sites relay positional signals during mouse brain development , 2000, Current Biology.

[8]  M. Ikura,et al.  Structural basis of calcium-induced E-cadherin rigidification and dimerization , 1996, Nature.

[9]  Jonathan A. Cooper,et al.  Reelin-induced tryosine phosphorylation of Disabled 1 during neuronal positioning , 1999 .

[10]  Xiang-Xi Xu,et al.  Disabled-2 inactivation is an early step in ovarian tumorigenicity , 1999, Oncogene.

[11]  Jonathan A. Cooper,et al.  Mouse disabled (mDab1): a Src binding protein implicated in neuronal development , 1997, The EMBO journal.

[12]  C. Kaiser,et al.  Methods in Yeast Genetics: A Cold Spring Harbor Laboratory Course Manual , 1994 .

[13]  T. Yagi,et al.  Proteins of the CNR Family Are Multiple Receptors for Reelin , 1999, Cell.

[14]  R. Heimark,et al.  Protocadherins: a large family of cadherin‐related molecules in central nervous system. , 1993, The EMBO journal.

[15]  Ramin Homayouni,et al.  Reelin Is a Ligand for Lipoprotein Receptors , 1999, Neuron.

[16]  M. Takeichi,et al.  Cadherins: a molecular family important in selective cell-cell adhesion. , 1990, Annual review of biochemistry.

[17]  Jonathan A. Cooper,et al.  Phospholipids of Transmembrane Glycoproteins and to Domain Binds to the Internalization Signals The Disabled 1 Phosphotyrosine-Binding , 1999 .

[18]  E. Robertis,et al.  The protocadherin PAPC establishes segmental boundaries during somitogenesis in Xenopus embryos , 2000, Current Biology.

[19]  F. Hoffmann,et al.  Dosage-sensitive modifiers of Drosophila abl tyrosine kinase function: prospero, a regulator of axonal outgrowth, and disabled, a novel tyrosine kinase substrate. , 1993, Genes & development.

[20]  E. D. De Robertis,et al.  Mouse paraxial protocadherin is expressed in trunk mesoderm and is not essential for mouse development , 2000, Genesis.

[21]  Jonathan A. Cooper,et al.  Neuronal position in the developing brain is regulated by mouse disabled-1 , 1997, Nature.

[22]  Shintaro T. Suzuki.  Structural and functional diversity of cadherin superfamily: Are new members of cadherin superfamily involved in signal transduction pathway? , 1996, Journal of cellular biochemistry.

[23]  T. Curran,et al.  Role of the reelin signaling pathway in central nervous system development. , 2001, Annual review of neuroscience.

[24]  S. Hirano,et al.  Expression of a Novel Protocadherin, OL-Protocadherin, in a Subset of Functional Systems of the Developing Mouse Brain , 1999, The Journal of Neuroscience.

[25]  F. van Roy,et al.  Phylogenetic analysis of the cadherin superfamily allows identification of six major subfamilies besides several solitary members. , 2000, Journal of molecular biology.

[26]  D. Hicklin,et al.  cDNA cloning, chromosomal mapping, and expression analysis of human VE-Cadherin-2 , 2000, Mammalian Genome.

[27]  Masahiko Watanabe,et al.  Diversity Revealed by a Novel Family of Cadherins Expressed in Neurons at a Synaptic Complex , 1998, Neuron.

[28]  D. Selkoe,et al.  Mutagenesis Identifies New Signals for β-Amyloid Precursor Protein Endocytosis, Turnover, and the Generation of Secreted Fragments, Including Aβ42* , 1999, The Journal of Biological Chemistry.

[29]  P. Worley,et al.  Arcadlin Is a Neural Activity-regulated Cadherin Involved in Long Term Potentiation* , 1999, The Journal of Biological Chemistry.

[30]  T. Yagi,et al.  Cadherin superfamily genes: functions, genomic organization, and neurologic diversity. , 2000, Genes & development.

[31]  T. Curran,et al.  A protein related to extracellular matrix proteins deleted in the mouse mutant reeler , 1995, Nature.

[32]  C. Rock,et al.  Cloning of a Novel Phosphoprotein Regulated by Colony-stimulating Factor 1 Shares a Domain with the Drosophiladisabled Gene Product (*) , 1995, The Journal of Biological Chemistry.

[33]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[34]  T. Curran,et al.  Identification of Reelin-induced Sites of Tyrosyl Phosphorylation on Disabled 1* , 2001, The Journal of Biological Chemistry.

[35]  J. Borg,et al.  Interaction of Cytosolic Adaptor Proteins with Neuronal Apolipoprotein E Receptors and the Amyloid Precursor Protein* , 1998, The Journal of Biological Chemistry.

[36]  S. Suzuki.  Recent progress in protocadherin research. , 2000, Experimental cell research.

[37]  F. Breviario,et al.  Identification of a Novel Cadherin (Vascular Endothelial Cadherin-2) Located at Intercellular Junctions in Endothelial Cells* , 1998, The Journal of Biological Chemistry.

[38]  Tom Maniatis,et al.  Large exons encoding multiple ectodomains are a characteristic feature of protocadherin genes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[39]  John Shelton,et al.  Reeler/Disabled-like Disruption of Neuronal Migration in Knockout Mice Lacking the VLDL Receptor and ApoE Receptor 2 , 1999, Cell.

[40]  Dan Goldowitz,et al.  Scrambler and yotari disrupt the disabled gene and produce a reeler -like phenotype in mice , 1997, Nature.

[41]  H. Kato,et al.  BH‐protocadherin‐c, a member of the cadherin superfamily, interacts with protein phosphatase 1 alpha through its intracellular domain , 1999, FEBS letters.

[42]  Ramin Homayouni,et al.  Disabled-1 Binds to the Cytoplasmic Domain of Amyloid Precursor-Like Protein 1 , 1999, The Journal of Neuroscience.

[43]  T. Maniatis,et al.  A Striking Organization of a Large Family of Human Neural Cadherin-like Cell Adhesion Genes , 1999, Cell.

[44]  Jonathan A. Cooper,et al.  Disabled‐2 Colocalizes with the LDLR in Clathrin‐Coated Pits and Interacts with AP‐2 , 2001, Traffic.

[45]  Joachim Herz,et al.  Direct Binding of Reelin to VLDL Receptor and ApoE Receptor 2 Induces Tyrosine Phosphorylation of Disabled-1 and Modulates Tau Phosphorylation , 1999, Neuron.

[46]  T. Bouwmeester,et al.  The role of paraxial protocadherin in selective adhesion and cell movements of the mesoderm during Xenopus gastrulation. , 1998, Development.