1. Yeast model of RNA binding proteins containing putative
prion-like domains in neurodegenerative diseases
Brittani M. Watkins. Amy E. Boncella, and Eric D. Ross
Department of Biochemistry & Molecular Biology, Colorado State University, Fort Collins, CO 80523
Background Methods
Results
Discussion
References
• Recently, several RNA-binding proteins
(RBPs) have been linked to
neurodegenerative diseases, including
amyotrophic lateral sclerosis (ALS) and
Alzheimer’s1,2
• Remarkably many of these RBPs,
including those already linked to
disease, contain prion-like domains
(PrLDs)1,2
• Algorithms created for yeast prions have
been used for determining PrLDs within
the human genome3
Purpose
• To begin evaluating the likelihood of
these proteins causing disease, we use
yeast to visualize their aggregation
ability
• Once we determine aggregation ability,
we can begin making algorithm-based
mutations to promote or inhibit
aggregation
• Give insight into the process behind
pathogenic aggregate formation
• If aggregation is occurring for the full-
length protein or its PrLD, then we will
visualize foci within the yeast cells
• Need to visualize the nucleus within Atx2
constructs
• Human homologs G3BP and TIA-1 had
similar phenotypes to Drosophila Ras and
Rox8 proteins, respectively2
• Drosophila homologs seem to mimic
human version within yeast
• Determining PrLD boundaries will be
important for modeling causative
mutations
• Plan to remove PrLDs from Atx2 full length
to get different perspective of PrLD
boundaries.
1. King, O. D., Gitler, A. D., and Shorter, J. (2012) The tip of the iceberg:
RNA-binding proteins with prion-like domains in neurodegenerative
disease. Brain Research 1462, 61-80
2. Couthouis, J., Hart, M. P., Shorter, J., DeJesus-Hernandez, M., Erion R.,
Oristano, R., Liu, A. X., Ramos, D., Jethava, N., Hosangadi, D., Epstein, J.,
Chiang, A., Diaz, Z., Nakaya, T., Ibrahim, F., Kim, H., Solski, J. A., Williams,
K. L., Mojsilovic-Petrovic, J., Ingre, C., Boylan, K., Graff- Radford, N. R.,
Dickson, D. W., Clay-Falcone, D., Elman, L., McCluskey, L., Greene, R.,
Kalb, G. A., Blair, I. P., King, O. D., Bonini, N. M., Deerlin, V. V.,
Rademakers, R., Mourelatos, Z., and Gitler, A. D. (2011) A yeast
functional screen predicts new candidate ALS disease genes. PNAS 108,
20881-20890
3. Cascarina, S. M., and Ross, E. D. (2014) Yeast prions and human prion-
like proteins: sequence features and prediction methods. Cell. Mol. Life
Sci. 71, 2047-2063
Acknowledgments
• A huge thank you to Eric D. Ross and all
the members of the Ross laboratory for
insightful discussion
PrLDs
Image adapted from Cascarina and Ross, 2014 3
Full length
Atx2’s PrLDs
• GFP with no fusion was used as a negative control
• Atx2 showed one large focus or possibly diffuse
within nucleus
• Ras cytoplasmic foci but also diffuse background
• Rox8 showed multiple cytoplasmic foci
• PrLD1 and PrLD3 were diffuse
• PrLD2 showed ring-like structures
• None of the PrLDs showed the exact same
phenotype as full length Atx2