Major Research Interests:

α-Synuclein and Parkinson’s disease; α-synuclein and melanoma; vesicle trafficking; molecular bases of disease.

α-Synuclein and Parkinson disease
The WITT group uses several organisms (yeast, mice and human cells) to study the mechanism of toxicity of the human Parkinson’s disease-associated protein a-synuclein (α-syn). α-Syn is an intrinsically unfolded protein of unknown function that is the main protein component of Lewy bodies, which are proteinaceous cytoplasmic inclusions in dopamine-producing neurons in individuals who suffer from PD.  High expression levels of a-syn or posttranslational modifications of the protein are thought to convert α-syn from a non-toxic protein into a toxic one.  There is increasing evidence that the toxic conformation of α-syn is a prion: it acts as a template or seed that converts non-infectious α-syn monomers into infectious oligomers.

Examples of ongoing projects include: 

(1) α-Syn intracellular trafficking and lipid dyshomeostasis:  In eukaryotic cells, wild-type a-syn transits through the endoplasmic reticulum and Golgi apparatus on route to the inner leaflet of the plasma membrane. In neurons, a-syn is thought to promote the fusion of presynaptic vesicles with the presynaptic membrane. We have found that the trafficking of a-syn through the ER is exquisitely sensitive to the level of certain cellular phospholipids, and we are investigating how changes in phospholipid homeostasis alter, and in some cases blocks, a-syn transit through the ER. This work has implications to the mechanism of Lewy body formation.

The figure below (Wang, 2014 PNAS) shows how a-syn aggregates form in yeast cells that lack the mitochondrial enzyme phosphatidylserine decarboxylase (Psd1). The role of this ancient enzyme vis-à-vis a-syn solubility/toxicity is an area of our focus.

(2) Viruses and Lewy body formation:  We have found that some viral proteins greatly accelerate the formation of a-syn fibrils in vitro. Usually, the half-time for fibril formation in vitro is 5 days, whereas in the presence of certain viral proteins the half-time is in the order of minutes! Using atomic force microscopy, we found that the fibrils formed in the absence of the viral protein are flat (A), whereas fibrils formed in the presence of the viral protein are helical (B). We are conducting a variety of kinetic and structural experiments to decipher how the viral peptides accelerate fibril formation.

(3)  a-Syn, iron homeostasis, retromer: Many studies over the last 10 years have found a link between a-syn and iron homeostasis. Red blood cells contain relatively high levels of a-syn as well as neurons. Because many of the genes involved in iron homeostasis in human cells are also found in yeast, we are using yeast to investigate the connection between a disruption of cellular iron homeostasis and a-syn toxicity. The findings from yeast are being tested in worms and human cells.

The figure below (Patel, 2018 Hum Mol Genet) shows how a-syn inhibits the Snx3-retromer-mediated recycling of Ftr1-Fet3. In yeast, Ftr1-Fet3 is a complex that imports iron into cells. The human orthologs of Fet3 are hephestin and ceruloplasmin.

(4)  a-Synuclein and melanoma: It is curious that individuals with melanoma (who live) have a 2-fold higher risk of being afflicted with PD than age-matched healthy controls. And it works the other way, that is, individuals with PD have a significantly higher risk of developing melanoma than age-matched individuals without PD. Melanocytes, like dopaminergic neurons, express α-syn. Melanocytes, like dopaminergic neurons, synthesize a pigment (melanin). Some of the most aggressive melanomas also express very high levels of α-syn, as if somehow α-syn promotes growth.  Experiments are underway using α-syn knockout cells to decipher the function of this interesting protein in melanoma.

Selected Publications

1. Patel, D,  Xu, C., Nagarajan, S., Liu, Z., Hemphill, W.O., Shi, R., Uversky, V.N., Caldwell, G.A., Caldwell, K.A., & Witt, S.N.  (2018)   Alpha -synuclein inhibits Snx3-retromer-mediated retrograde recycling of iron transporters in S. cerevisiae and C. elegans models of Parkinson’s disease.   Human Molecular Genetics 27(9), 1514–1532
2. Wang, S., Zhang, S., Liou, L.-C., Ren, Q., Zhang, Z., Caldwell, G.A., Caldwell, K.A., & Witt, S.N. (2014) Phosphatidylethanolamine deficiency disrupts α-synuclein homeostasis in yeast and worm models of Parkinson disease. Proceedings of the National Academy of Sciences U S A 111(38): E3976–E3985
3. Witt, S.N. (2013). Molecular chaperones, alpha-synuclein and neurodegeneration.  Molecular Neurobiology 47, 552-560.   
4. Wang, S., Xu, B., Liou, L.-C., Ren, Q., Huang, S., Luo, Y.,  Zhang, Z. & Witt, S.N. (2012). α-Synuclein disrupts stress signaling by inhibiting polo-like kinase Cdc5/Plk2.  Proceedings of the National Academy of Sciences U S A 109, 16119-16124.
5. Liu, X.,  Lee, Y.L.,  Liou, L.-C.,  Ren, Q., Zhang, Z., Wang, S. & Witt, S.N. (2011). Alpha-synuclein protects cells from hydroxyurea-induced replication stress in a yeast model of Parkinson’s disease. Human  Molecular Genetics  20, 3401-3414.
6. Lee,  Y.J., Wang, S.,  Slone, S.R., Yacoubian, T.A. & Witt, S.N. (2011). Defects in very long chain fatty acid synthesis enhance alpha-synuclein toxicity in a yeast model of Parkinson’s disease. PLoS ONE  6 (1):e15946
7. Witt, S.N. (2010). Hsp70 molecular chaperones and Parkinson’s disease. Biopolymers  93, 218-28.
8. Liang, J.,  Clark-Dixon, C., Wang, S., Flower, T.R., Williams-Hart, T., Zweig, R.,  Robinson, L.C.,  Tatchell, K. & Witt, S.N. (2008). Novel suppressors of alpha-synuclein toxicity identified using yeast.  Human  Molecular Genetics  17, 3784-3795.
9. Flower, T.R., Clark-Dixon, C., Metoyer, C., Yang, H., Shi, R., Zhang, Z. & Witt, S.N. (2007). YGR198w (YPP1) targets A30P alpha-synuclein to the vacuole for degradation.  Journal of Cell Biology 177, 1091-1104.
10. Flower, T.R., Chesnokova, L.S., Froelich, C.A., Dixon, C. & Witt, S.N. (2005). Heat shock prevents alpha synuclein-triggered apoptosis in a yeast model of Parkinson’s disease. Journal of Molecular Biology 351, 1081-1100. 

Post-doctoral Fellows

We are currently accepting applications for Post-doctoral Fellows.

Graduate Students

We are currently accepting applications for Graduate students interested in conducting research in the Witt lab.

Undergraduate Research Assistants

We are currently accepting applications from undergraduates. 

Medical Students, Residents, and Fellows

The Witt laboratory has a number of research projects available for any Medical Students, Residents, and Fellows interested. Contact the lab for more information.