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Rotin

Research Interests

• Protein Interactions
• Proteomics
• Signal transduction
• Tyrosine Phosphatases
• Ubiquitination
• Epithelial Na channel
• Genetics of hypertension
• Ras activators

Research Activities

Protein: protein interaction domains and ubiquitination:
My laboratory has been studying the role of protein and domains which interact and regulate the epithelial Na channel (ENaC). We identified the ubiquitin ligase (Hect-domain family) Nedd4 as a protein that binds the PY motifs of ENaC via its WW domain and suppresses the number of channels at the cell surface by ubiquitination and degradation; these PY motifs are mutated in Liddle syndrome patients, a hereditary form of hypertension caused by hyperactivity of the channel in the kidney. The C2 domain of Nedd4 mediates its apical membrane localization via a calcium-dependent interaction with the apical raft-associated protein annexin XIIIb. Nedd4 also has other substrates, including the novel cAMP-dependent Ras activator CNrasGEF.

LAR family protein tyrosine phosphatases (PTPs):
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 severe neuroendocrine and neuronal defects. The catalytic domains of LAR family members interact with each other to inhibit enzymatic activity. We are now attempting to identify the substrate(s) of PTPsigma and the signaling pathway it is involved in.

Future Research Interests

Protein: protein interaction domains and ubiquitination: We are planning to utilize proteomic approaches to understand global regulation of substrates of ubiquitin ligases such as Hect-E3s (eg Nedd4 or Nedd4-like proteins), while further characterizing the substrates we have already identified (ENaC, CNrasGEF, etc).

LAR family protein tyrosine phosphatases (PTPs): Once we identify the substrate(s) of PTPsigma, we will study its role s in signaling and cell biology of neuronal cells including its role in axon guidance.

Publications

Kus B, Rouleau N, Chan D, Sun W, Gajadhar A, Lenk A, Lee T, Edwards A, Bosse R, ROTIN D. A high throughput screen to identify substrates for the Ubiquitin ligase Rsp5. J Biol Chem. 280(33):29470-8, 2005.

Rougier JS, Van Bemmelen M, O'Brien MC, Jespersen T, Gavillet B, Apotheloz F, Cordonier S, Staub S, ROTIN D, Abriel H. Molecular determinants of voltage gated sodium channels regulation by Nedd4/Nedd4-like proteins. Am J. Physiol (cell), C692-701,2005.

Ishikawa T, Jiang C, Stutts J, Marunaka Y, ROTIN D. Regulation of the epithelial Na channel (rENaC) by cytosolic ATP. J. Biol. Chem. 278:38276-86, 2003.

Henry P, Kanelis V, Kim B, O'Brien MC, Forman-Kay J, Schild L, and ROTIN D. Affinity and specificity of interactions between Nedd4 and ENaC. J. Biol. Chem. 278:20019-28, 2003.

Vecchione A, Marchese A, Henry P, ROTIN D, Morrione A. Grb10/Nedd4 complex regulates ligand-induced ubiquitination and stability of the Insulin-like Growth Factor 1 Receptor. Mol. Cell Biol. 23:3363-3372, 2003.

Batt J, Cutz E, Fladd C, ROTIN D. Apparent normal lung architecture in PTPsigma deficient mice. Am. J. Physiol, 284:L214-223, 2003.

Pak Y, Pham N, ROTIN D. Direct binding of the b1 adrenergic receptor to the cAMP dependent guanine nucleotide exchange factor CNrasGEF leads to Ras activation. Mol. Cell Biol. 22:7942-7952, 2002.

Myat A* / Henry P*, McCabe V, Flintoft L, ROTIN D, Tear G. Nedd4, a ubiquitin ligase, is recruited by commissureless to control cell surface levels of the roundabout receptor. Neuron 35:447-459, 2002.

Meathrel K, Adamek T, Batt J, ROTIN D, Doering L. PTPs deficient mice show spinal cord abnormalities and aberrant cytoarchitecture in the cortex, hippocampus and corpus callosum. J. Neurosci. Res. 70:24-35, 2002.

McLean J, Batt J, Doering L, ROTIN D, 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.

Hanwell D, Ishikawa T, Saleki R, ROTIN D. Trafficking and cell surface stability of ENaC expressed in epithelial MDCK cells. J. Biol. Chem., 277:9772-9779, 2002.

Batt J, Asa S, Fladd C, ROTIN D. Pituitary, pancreatic and gut neuroendocrine defects in PTPsigma deficient mice. Mol. Endocrinol., 16:155-169, 2002.

Pham N, ROTIN D. Nedd4 regulates ubiquitination and stability of the guanine nucleotide exchange factor CNrasGEF. J. Biol. Chem. 276:46995-47003, 2001.

Kanelis V, ROTIN D, Forman-Kay J. Solution structure of a Nedd4 WW domain -bENaC peptide complex. Nature Struct. Biol. 8:407-412, 2001.

Kanelis V, ROTIN D, Forman-Kay, J. Solution structure of a Nedd4 WW domain - ENaC peptide complex. Nature Struct. Biol. 8:407-412, 2001.

Staub O, Abriel H, Plant P, Ishikawa T, Saleki R, Schild L, ROTIN D. Regulation of ENaC by Nedd4 and by ubiquitination. Kidney Internat. 57:809-815, 2000.

ROTIN D, Staub O, Haguenauer-Tsapis R. Ubiquitination and endocytosis of plasma membrane proteins; Role of Nedd4/Rsp5 family of ubiquitin protein ligases. J. Membrane Biology. 176:1-17, 2000.

Pham N, Cheglakov I, Koch A, de-Hoog C, Moran MF and ROTIN D. The guanine nucleotide exchange factor CNrasGEF activates Ras in response to cAMP and cGMP. Curr. Biol. 10:555-558, 2000.

Plant P, Lafont F, Lecat S, Verkade P, Simons K, 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.

Winberg G, Matskova L, Chen F, Plant P, ROTIN D, Gish G, Ingham R, Pawson T. LMP2A protein of Epstein-Barr virus binds WW domain E3 protein ubiquitin ligases that ubiquitinate B cell tyrosine kinases. Mol. Cell Biol. 20:8526-8535, 2000.

Morrione A, Plant P, Valentinis B, Staub O, Kumar S, ROTIN D, Baserga R. mGrb10 interacts with Nedd4. J. Biol. Chem. 274:24094-24099, 1999.

Wallace MJ, Batt J, Fladd C, Henderson JT, Skarnes W, 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, Staub O. Defective regulation of the epithelial Na+ channel (ENaC) by Nedd4 in Liddle's Syndrome. J. Clin. Invest., 103:667-673, 1999.

Ishikawa T, Marunaka Y, ROTIN D. Electrophysiological characterization of αβγENaC expressed in MDCK cells: Role of Na+ and Ca2+. J. Gen. Physiol. 111:825-846, 1998.

Wallace M, Fladd C, Batt J, ROTIN D. The second catalytic domain of the tyrosine phosphatase PTPδ binds to and inhibits the first catalytic domain of PTPσ. Mol. Cell. Biol.,18:2608-2616, 1998.

Plant P, Yeger H, Staub O, Howard P, ROTIN D. The C2 domain of the ubiquitin protein ligase Nedd4 mediates Ca2+-dependent plasma membrane localization. J. Biol. Chem., 272:32329-32377, 1997.

Staub O, Gautschi I, Ishikawa T, Breitschopf K, Ciechanover A, Schild L, ROTIN D. Regulation of stability and function of the epithelial Na channel (ENaC) by ubiquitination. EMBO J. 16:6325-6336,1997.

ROTIN D. WW (WWP) Domains: From structure to function. In "Modular Domains in Signal Transduction", Current Topics in Microbiology and Immunology (Editor: T. Pawson). Vol 228, pp.115-133, 1997.

Staub O, Dho S, Henry P, Correa J, Ishikawa T, McGlade J, 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.

Staub O, ROTIN D. WW Domains. Structure (Curr. Biol.),4:495-499, 1996.