Deposition of aggregates of misfolded protein into intracellular inclusion bodies is a prominent cytopathological feature of nearly every known neurodegenerative disease. Despite mounting genetic and biochemical evidence linking protein aggregation to pathogenesis in these and other diseases, it is unclear bow -or indeed whether- protein aggregation and inclusion body formation are primary toxic events or cytoprotective responses. My lab has recently described a general pathway by which aggregated proteins in mammalian cells are collected into specialized inclusion bodies called aggresomes (AG). The studies described in this proposal are intended to test the hypothesis that delivery of protein aggregates to AG is a specific, microtubule-dependent transport process which facilitates the neutralization and elimination of potentially toxic gene products. Towards this end, three specific aims are proposed.
The first aim will use biochemical and biophysical techniques to study the cellular mechanism of AG formation to identify transport intermediates in AG formation. These intermediates will be subject to extensive biochemical, biophysical and structural characterization.
The second aim of the proposed research will be to reconstitute AG formation in a cell-free system in order to identify the cytoplasmic components required for retrograde transport of protein aggregates on microtubule tracks. Finally, the last aim will investigate the role of retrograde transport in the neutralization and elimination of protein aggregates. These last studies will specifically test the hypothesis that retrograde transport of aggregated protein is linked to the lysosomal/autophagic pathway of protein degradation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
1R01NS042842-01A1
Application #
6545585
Study Section
Special Emphasis Panel (ZRG1-MDCN-2 (01))
Program Officer
Murphy, Diane
Project Start
2002-06-01
Project End
2007-05-31
Budget Start
2002-06-01
Budget End
2003-05-31
Support Year
1
Fiscal Year
2002
Total Cost
$335,588
Indirect Cost
Name
Stanford University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Pearce, Margaret M P; Kopito, Ron R (2018) Prion-Like Characteristics of Polyglutamine-Containing Proteins. Cold Spring Harb Perspect Med 8:
Bersuker, Kirill; Brandeis, Michael; Kopito, Ron R (2016) Protein misfolding specifies recruitment to cytoplasmic inclusion bodies. J Cell Biol 213:229-41
Pearce, Margaret M P; Spartz, Ellen J; Hong, Weizhe et al. (2015) Prion-like transmission of neuronal huntingtin aggregates to phagocytic glia in the Drosophila brain. Nat Commun 6:6768
Bersuker, Kirill; Hipp, Mark S; Calamini, Barbara et al. (2013) Heat shock response activation exacerbates inclusion body formation in a cellular model of Huntington disease. J Biol Chem 288:23633-8
Streets, Aaron M; Sourigues, Yannick; Kopito, Ron R et al. (2013) Simultaneous measurement of amyloid fibril formation by dynamic light scattering and fluorescence reveals complex aggregation kinetics. PLoS One 8:e54541
Trevino, R Sean; Lauckner, Jane E; Sourigues, Yannick et al. (2012) Fibrillar structure and charge determine the interaction of polyglutamine protein aggregates with the cell surface. J Biol Chem 287:29722-8
Hipp, Mark S; Patel, Chetan N; Bersuker, Kirill et al. (2012) Indirect inhibition of 26S proteasome activity in a cellular model of Huntington's disease. J Cell Biol 196:573-87
Hipp, Mark S; Bersuker, Kirill; Kopito, Ron R (2012) Live-cell imaging of ubiquitin-proteasome system function. Methods Mol Biol 832:463-72
Ryu, Kwon-Yul; Park, Hyejin; Rossi, Derrick J et al. (2012) Perturbation of the hematopoietic system during embryonic liver development due to disruption of polyubiquitin gene Ubc in mice. PLoS One 7:e32956
Riley, Brigit E; Kaiser, Stephen E; Kopito, Ron R (2011) Autophagy inhibition engages Nrf2-p62 Ub-associated signaling. Autophagy 7:338-40

Showing the most recent 10 out of 23 publications