Daniela Rotin , PhD
University of Toronto
Canada Research Chair
Biochemistry and Signal Transduction
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- Protein interactions
- Signal transduction
- Ion channels (ENaC, CFTR)
- Tyrosine phosphatases
- Human diseases (Cystic fibrosis, IBD, Cancer)
Biochemistry and Function of the Nedd4 Family of Ubiquitin Ligases: The major focus of my laboratory has been the ubiquitin system, particularly the Nedd4 family of E3 ubiquitin ligases, comprised of a C2-WW(n)-HECT domain architecture. We are studying the biochemistry, structure and function of these E3 ligases, as well as develop methodologies to globally identify their substrates. We also study in more detail how they regulate several of these substrates (eg. ENaC, LAPTM5, FGFR1). More recently, we have been using mouse knockout and other animal models of Nedd4 family members to decipher their biological function in vivo.
Two additional projects in the lab focus on (a) LAR family protein tyrosine phosphatases (PTPs), and on (b) rescue of mutant CFTR: In (a) we have been characterizing the LAR family protein PTPsigma both biochemically and developmentally. We knocked out the PTPsigma gene and showed that the mice develop neuroendocrine and neuronal defects. They also exhibit intestinal abnormalities. We recently identified N and E cadherin as in vivo substrates for PTPsigma in the brain and intestine, respectively. We then showed that polymorphism in the PTPsigma gene is associated with ulcerative colitis, and polymorphism in its substrate E-cadherin is associated with Crohn's disease, two IBD illnesses. In (b), we have developed screens to identify proteins and small molecules/drugs that rescue the major mutation in cystic fibrosis, the deltaF508-CFTR.
Future Research Interests
We are interested in further studying the biological functions of the Nedd4 family of E3 ubiquitin ligases and their substrates using animal models, to understand their roles in normal development, in disease and in tissue regeneration.
We are also interested in analyzing the role of the tyrosine phosphatase PTPsigma and its substrates in the regulation of intestinal epithelial integrity and in IBD.
As well, we will be investigating the mechanisms by which our recently identified correctors of the major mutation in cystic fibrosis carry out their functions.
Trzcinska-Daneluti A., Nguyen L., Jiang C., Fladd C., Al-Awar, R. and ROTIN D. Kinase inhibitors identified in kinome suppressor screens correct the trafficking defect of ∆F508-CFTR. Mol Cell Proteom. 11:745-757, 2012
Persaud A., Alberts P, Hayes H, Guettler S, Clarke I, Sicheri F, Dirks P, Ciruna B and ROTIN D. Nedd4-1 Binds and Ubiquitylates Activated FGFR1 to Control its Endocytosis and Function. EMBO J. 30:3259-3272, 2011
Kimura T, Kawabe H, Jiang C, Zhang W, Xiang YY, Lu C, Salter MW, Brose N, Lu W-Y and ROTIN D. Deletion of the ubiquitin ligase Nedd4L in lung epithelia causes cystic fibrosis-like disease. Proc.Natl.Acad.Sci. USA, 108: 3216-3221,2011
ROTIN D. and Kumar S. Physiological functions of the HECT family of ubiquitin ligases. Nature Rev. Mol. Cell. Biol. 10:398-409, 2009
Persaud A., Amsen E., Xiong X, Wasmuth J, Saadon Z, Fladd C, Parkinson J. and ROTIN D. Comparison of substrate specificity between the ubiquitin ligases Nedd4 and Nedd4-2 using proteome arrays. Mol Syst Biol. 5:333, 2009.
Trzcinska-Daneluti A., Ly, D., Huynh L., Jiang C., Fladd C and ROTIN D. High content functional screen to identify proteins that correct F508del-CFTR function. Mol.Cell.Proteom.8:780-790, 2009
Fouladkou F., Landry T., Kawabe H., Lu, C., Brose N., Stambolic V and ROTIN D. The Ubiquitin ligase Nedd4-1 is dispensable for the regulation of PTEN stability, localization or activity. Proc. Natl. Acad. Sci, USA, 105:8585-8590, 2008
Wiesner S., Ogunjimi A., Wang HR., ROTIN D., Sicheri F., Wrana J., Forman-Kay J. Auto-inhibition of the Hect-type ubiquitin ligase Smurf2 through its C2 domain. Cell, 130:651-62, 2007.
Muise A., Walters T., Wine E., Griffiths A., Duerr R., Turner D., Regueiro M., Ngan BY., Sherman P., Siverberg M. and ROTIN D. Protein Tyrosine Phosphatase sigma is associated with Ulcerative Colitis. Curr. Biol. 17:1212-1218, 2007.
Lu C., Pribanic S., Gaulis A. Jiang C. and ROTIN D. The PY motif of ENaC, mutated in Liddle syndrome, regulate channel internalization, sorting and mobilization from a sub-apical pool. Traffic, 8:1246-1264, 2007.
Gupta R., Kus B., Fladd C., Wasmuth J., Tonikian R., Krogan N., Sidhu S., Parkinson J. and ROTIN D. High throughput ubiquitination screen using protein microarrays for the comprehensive identification of Rsp5 substrates in yeast. Mol Syst Biol. 3: (116) 1-12, 2007.
Siu, R., Fladd C. and ROTIN D. N-cadherin is an in vivo substrate for PTPsigma and participates in PTPsigma-mediated inhibition of axon growth Mol. Cell Biol. 27:208-219, 2007.
Pak Y., Glowacka W., Bruce C., Pham N. and ROTIN D. Transport of LAPTM5 to lysosomes requires association with the ubiquitin ligase Nedd4 but not LAPTM5 ubiquitination. J. Cell Biol. 175:61-645, 2006.
McLean J., Batt J., Doering L., ROTIN D. and Bain JR. Enhanced rate of regeneration and directional errors following sciatic nerve injury in receptor protein tyrosine phosphatase sigma knockout mice. J. Neurosci. 22:5481-5491, 2002
Kanelis V., ROTIN D*. and Forman-Kay, J*. Solution structure of a Nedd4 WW domain -ENaC peptide complex. Nature Struct. Biol. 8:407-412, 2001. (* co-PIs)
Plant P., Lafont F., Lecat S., Verkade P., Simons K. and ROTIN D. Apical membrane localization of Nedd4 is mediated by an association of its C2 domain with Annexin XIIIb. J. Cell Biol. 149:1473-1483, 2000.
Wallace MJ., Batt J., Fladd C., Henderson JT., Skarnes W. and ROTIN D. Neuronal defects and posterior pituitary hypoplasia in mice lacking the receptor tyrosine phosphatase PTPσ. Nature Genet.,21:334-338, 1999.
Abriel H., Loffing J., Rebhun JF Pratt H, Schild L., Horisberger J-D, ROTIN D. and Staub O. Defective regulation of the epithelial Na+ channel (ENaC) by Nedd4 in Liddle’s Syndrome. J. Clin. Invest., 103:667-673, 1999.
Staub O, Gautschi I., Ishikawa T., Breitschopf K., Ciechanover A., Schild L and ROTIN D. Regulation of stability and function of the epithelial Na channel (ENaC) by ubiquitination. EMBO J. 16:6325-6336,1997.
Staub O, Dho S, Henry P, Correa J., Ishikawa T., McGlade J, and ROTIN D. WW domains of Nedd4 bind to the proline rich PY motifs in the epithelial Na channel deleted in Liddle’s syndrome. EMBO J, 15:2371-2380, 1996.