Faculty of Medicine / University of Toronto  

Signal Transduction in Normal Development and Disease

My research is directed towards understanding the regulation of protein interactions and signal-transduction pathways that control normal cellular function and human disease.  Projects in the lab are currently focused within three main themes:

Regulation of  signal transduction by adaptor proteins
My colleagues and I have focused the identification and functional characterization of intracellular adapter proteins that function to integrate, localize, and down regulate signal-transduction cascades.  My laboratory has cloned and characterized a number of new adapters such as Gads and SLAP/SLAP2, which function in both growth factor and antigen-receptor signaling, as well as the adaptors NUMB, EHD and Lulu/Ymo1, which regulate the activity of trans-membrane receptors that determine cell fate and polarity during development. Currently,we are working on the functional characterization of these molecules, and their associated protein networks.

Ubiquitin dependent regulation of signal transduction and cell polarity
Our lab is studying the role of ubiquitin dependent regulation of signaling pathways.  We have identified a family of novel E3 ubiquitin ligases, including LNX, and our current focus is to discover the role of LNX activity in cell fate determination and the establishment of cell polarity during embryonic development.  Our lab has also recently discovered a novel ubiquitin ligase, that functions in receptor tyrosine kinase trafficking. We are currently developing  high-throughput assays for LNX and RNF126 substrates and binding partners.

Development of high-throughput functional protein assays
As a co-director of  Signaling and Degradation Network (SIDNET, www.sickkids.ca/sidnet),  at the Hospital for Sick Children, my lab is also involved in the development of high throughput assays of signal transduction and protein degradation pathways using robotics, protein arrays and high throughput detection and imaging systems. The SIDNET platform capabilities will continue to be expanded to include disease specific sets of cDNAs in multi-purpose expression formats, and shRNA libraries.

More information can be found at http://www.sickkids.ca/mcglade

Graduate Students:

• Jonathan Krieger
• Larissa Liontos
• Brittany Prevost
• Christopher Smith
• Cheryl Wolting

Selected References:

Link to Pubmed Publications

Loreto, M.P., McGlade, C.J. (2003) Cloning and characterisation of human Src-like adaptor protein 2 and a novel splice isoform, SLAP-2-v. Oncogene,  22: 266-273.

Liu, Q., Berry, D., Nash P., Pawson, T., McGlade, CJ., Li, S-C. (2003) Structural Basis for Specific Binding of the Gads SH3 Domain to an RxxK Motif-Containing SLP-76 Peptide. A Novel Mode of Peptide Recognition.  Molecular Cell, 11:471-481.

McGill, M.A. and McGlade C.J. (2003) Mammalian Numb proteins inhibit Notch nuclear activity and promote Notch ubiquitination. J. Biol. Chem.  278:23196-203.

J. Nie,  S.C. Li and C. J. McGlade.  (2004) A novel PTB-PDZ domain interaction mediates isoform-specific ubiquitylation of mammalian Numb.  J. Biol. Chem. 279:20807-20815. 

Smith, C.A., Dho, S.E., and C.J. McGlade. (2004) The cell fate determinant Numb interacts with EHD/Rme-1 family proteins and has a role in endocytic recycling.   Mol. Biol. Cell.15: 3698-3708.

Young, P., Nie, J., Wang, X., McGlade, CJ., Rich, MM., G. Feng. (2005) LNX1 is a perisynaptic  Schwann cell specific E3 ubiquitin ligase that interacts with ErbB2. Mol. Cell. Neuroscience, 30(2):238-248.
 
Simoncic,  D.P., Bourdeau, A., Lee-Loy, A., Rohrschneider, L.R., Tremblay, M.L., Stanley, E.R. and C.J. McGlade. (2006) T-cell Protein Tyrosine  Phosphatase (Tcptp) is a negative regulator of CSF-1 signaling and mononuclear  phagocyte development.  Mol. Cell Biol.  26:4149-4160.

Laprise,  P. Beronja,  S., Silva-Gagliardi, N., Pellikka, M., Jensen, A., McGlade, C.J. and U. Tepass. (2006) The FERM protein Yurt is a negative regulatory component of the Crumbs complex that controls epithelial polarity and apical membrane size.  Dev. Cell, 11(3):363-374.

Dho, S.E.,  Siderovski, D.P.,  Trejo, J., and C. J. McGlade (2006) Dynamic regulation of mammalian numb by G protein-coupled receptors and protein kinase C activation. Mol. Biol. Cell.17:4142-55

Wolting , C., McGlade, CJ., and D. Tritchler (2006) Cluster analysis of protein array results via similarity of gene ontology annotation.  BMC Bioinformatics, 7:338.
 
Smith,  CA.,  Lau, K., Rahmani, Z., Dho, SE, Brothers, G., She, Y., Bonneil, E., Thibault, P., Berry, DM, Le Borgne. R., Shweisguth, F., and C.J. McGlade (2007) aPKC-mediated phosphorylation regulates asymmetric membrane localization of the cell fate determinant Numb. EMBO J. 26:468-80.

Seet, B.T., Berry, D.M., Maltzman, J., Shabason, J., Raina, M., Koretzky, G.A., McGlade, C.J., and T. Pawson. (2007) TCR signaling requires a high affinity interaction between Gads C-SH3 domain and the SLAP-76 RXXK motif. EMBO J 26:678-689.

Lee, J.D., Silva-Gagliardi, N.F., Tepass, U., McGlade,  C..J., and K.V. Anderson. (2007) the FERM protein Epb4.1l5 is required for organization of the neural plate and the epithelial-mesenchymal transition at the primitive streak of the mouse. Development. 134(11):2007-16

Pakuts, B. Debonneville, C., Lionto, L., Loreto, M.P., and C.J. McGlade (2007) SLAP-2 mediates c-Cbl dependent down regulation of Colony Stimulating Factor-1 Receptor  signaling.  J. Biol. Chem. 282:17953-63.