Investigating Proteolytic Processing of Ataxin 2, a Neurodegenerative Disease Associated Protein
dc.contributor.advisor | Patrick Emery | en_US |
dc.contributor.author | Chitre, Monika | |
dc.date.accessioned | 2022-09-20T14:59:45Z | |
dc.date.available | 2022-09-20T14:59:45Z | |
dc.date.issued | 2022-08-08 | |
dc.identifier.doi | 10.13028/py2x-sn09 | en_US |
dc.identifier.uri | http://hdl.handle.net/20.500.14038/51128 | |
dc.description.abstract | Ataxin 2 (ATXN2) is a ubiquitously expressed mRNA binding protein involved in the development and progression of spinocerebellar ataxia 2 (SCA2) and amyotrophic lateral sclerosis (ALS). In the context of both neurodegenerative diseases, its N-terminal polyglutamine (polyQ) domain is mutated and expanded in length. Several other polyQ proteins, such as huntingtin (Htt), ataxin 3 (ATXN3), and ataxin 7 (ATXN7), undergo proteolytic processing that produces toxic fragments containing their polyQ domains. Investigating how ATXN2 is regulated by proteolysis is hindered by the lack of available molecular biological tools such as N-terminal ATXN2 antibodies to target and analyze the endogenous N-terminus of ATXN2. To circumvent this challenge, I developed a transient overexpression model of N-terminally tagged ATXN2 in HEK293E cells. Here, I demonstrate that both wild-type and mutant ATXN2 are targets of N-terminal proteolysis. I confirmed that ATXN2 produces an independent polyQ cleavage fragment like other polyQ proteins through basic molecular biology approaches such as Western blotting and immunoprecipitation. Additionally, I identified the specific region that is both necessary and sufficient for cleavage to occur via deletion mapping with multiple truncated ATXN2 mutants and reporter constructs. Further definition of ATXN2 as a target of proteolytic cleavage aligns it with other neurodegenerative polyQ proteins, and proteolysis is currently a less explored avenue of research for ATXN2-related disease development, progression, and therapeutic modalities. This work reveals a novel site that directs cleavage of ATXN2 and provides a potential avenue of investigation for how ATXN2 posttranslational modifications contribute to the progression of SCA2 and ALS. | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | UMass Chan Medical School | en_US |
dc.rights | © 2022 Chitre. | en_US |
dc.rights.uri | All Rights Reserved | en_US |
dc.subject | neurodegeneration | en_US |
dc.subject | proteolysis | en_US |
dc.subject | ataxin 2 | en_US |
dc.title | Investigating Proteolytic Processing of Ataxin 2, a Neurodegenerative Disease Associated Protein | en_US |
dc.type | Doctoral Dissertation | en_US |
atmire.contributor.authoremail | monika.chitre@gmail.com | en_US |
dc.contributor.department | Neurobiology | en_US |
dc.contributor.department | Emery Lab | |
dc.contributor.student | Monika Chitre | en_US |
dc.description.thesisprogram | Interdisciplinary | en_US |