The lab studies cell polarity which is required for virtually all aspects of cell behavior including cell morphology, cell motility, and cell identity arising from asymmetric divisions (e.g., stem cells). In particular we are focused on how extracellular signals control metastatic behavior in melanoma by regulating cell polarity. Wnt5a is a non-canonical Wnt ligand that regulates cell polarity in developmental systems. We have found that Wnt5a induces the formation of a novel polarized structure (WRAMP structure) in melanoma cells that mediates membrane retraction and directional cell movement. We are using biochemistry and proteomics to identify the components of the WRAMP structure and live cell imaging to observe the temporal assembly of the components. Future work will determine the in vivo function of the WRAMP structure and regulated cell polarity during metastasis.
Our research is focused on the regulation of cell polarity in cancer and how it determines cell behavior during the transition from a non-metastatic to a metastatic state. We previously identified a novel polarized structure composed of an IgG family cell adhesion molecule named melanoma cell adhesion molecule (MCAM) forms in response to Wnt5a. We named this structure the "WRAMP (Wnt5a-induced, receptor, actin and myosin II polarity) structure", based on its recruitment of the MCAM cell adhesion receptor, Frizzled receptors, and an overlapping polarized complex of cytoskeletal proteins including F-actin and myosin IIB. Formation of the WRAMP structure led to rapid retraction of adjacent cell membrane, and moved the nucleus towards the side of the cell opposite from the WRAMP structure, leading to directional cell movement (Witze et al. Science 2008). Proteomic analysis has identified many proteins that localize to the WRAMP structure and many of these proteins are involved in cytoskeletal reorganization and/or membrane trafficking. This may suggest that Wnt signaling regulates polarity through the polarized trafficking of membrane compartments and proteins. 3D electron microscopy together with live imaging revealed that the endoplasmic reticulum also localizes to the WRAMP structure. This movement of the ER coincided with a localized elevation of calcium levels specifically in the WRAMP structure. These studies reveal a novel mechanism for the polarized distribution of cell structures that regulate calcium signaling spatially in response to Wnt5a in migrating cells.
01. Bloss TA, Witze ES, Rothman JH. Suppression of CED-3-independent apoptosis by mitochondrial betaNAC in Caenorhabditis elegans. Nature. 424(6952):1066-71. 2003.
02. Witze ES, Old WM, Resing KA, Ahn NG. Mapping protein post-translational modifications with mass spectrometry. Nat Methods. 4(10):798-806. 2007.
03. Witze ES, Litman ES, Argast GM, Moon RT, Ahn NG. Wnt5a control of cell polarity and directional movement by polarized redistribution of adhesion receptors. Science. 320(5874):365-9. 2008.
04. Witze ES, Field ED, Hunt DF, Rothman JH. C. elegans pur alpha, an activator of end-1, synergizes with the Wnt pathway to specify endoderm. Dev Biol. 327(1):12-23. 2009.
05. Old WM, Shabb JB, Houel S, Wang H, Couts KL, Yen CY, Litman ES, Croy CH, Meyer-Arendt K, Miranda JG, Brown RA, Witze ES, Schweppe RE, Resing KA, Ahn NG. Functional proteomics identifies targets of phosphorylation by B-Raf signaling in melanoma. Mol Cell. 34(1):115-31. 2009.
06. Kabuyama Y, Litman ES, Templeton PD, Metzner SI, Witze ES, Argast GM, Langer SJ, Polvinen K, Shellman Y, Chan D, Shabb JB, Fitzpatrick JE, Resing KA, Sousa MC, Ahn NG. A mediator of Rho-dependent invasion moonlights as a methionine salvage enzyme. Mol Cell Proteomics. 8(10):2308-20. 2009.
07. Sayers CM, Papandreou I, Guttmann DM, Maas NL, Diehl JA, Witze ES, Koong AC, Koumenis C. Identification and Characterization of a Potent Activator of P53-Independent Cellular Senescence Via a Small Molecule Screen for Modifiers of the Integrated Stress Response. Mol Pharmacol. 83(3):594-604. 2013.
08. Witze ES, Connacher MK, Houel S, Schwartz MP, Morphew MK, Reid L, Sacks DB, Anseth KS, Ahn NG. Wnt5a directs polarized calcium gradients by recruiting cortical endoplasmic reticulum to the cell trailing edge. Dev Cell. 26(6):645-57. 2013.
09. Londoño Gentile T, Lu C, Lodato PM, Tse S, Olejniczak SH, Witze ES, Thompson CB, Wellen KE DNMT1 is regulated by ATP-citrate lyase and maintains methylation patterns during adipocyte differentiation. Mol Cell Biol. 33(19):3864-78. 2013.
10. Wang W, Terkowski S, Ekaireb R, Runkle K, Putzke A, Witze E Wnt5a induced protein depalmitoylation regulates developmental cell movement and tumor cell invasion. Oncogenesis (4) 2015.
11. Wang W, Snyder N, Worth AJ, Blair IA, Witze ES Regulation of lipid synthesis by the RNA helicase Mov10 controls Wnt5a production. JBC 290(25): 15707-16. 2015
12. Runkle K, Kharbanda A, Stypulkowski E, Cao X-J, Wang W, Garcia, B, Witze E Inhibition of DHHC20 mediated EGFR palmitoylation creates a dependence on EGFR signaling. Molecular Cell (In Press).
Akriti Kharbanda - Graduate Student
Ewa Stypulkowski - Graduate Student
Katelyn Carlin - Administrative Coordinator
Rochelle Sadeghi - Graduate Student
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