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Cell biological approaches to investigate polyglutamine-expanded AR metabolism. | LitMetric

Cell biological approaches to investigate polyglutamine-expanded AR metabolism.

Methods Mol Biol

Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, USA.

Published: December 2013

AI Article Synopsis

Article Abstract

Spinal and bulbar muscular atrophy (SBMA) is a late-onset neurodegenerative disease caused by a polyglutamine expansion in the androgen receptor (AR). In vivo and in vitro studies have suggested that some steps of normal AR function and metabolism, such as hormone binding and nuclear translocation of the AR, are necessary for toxicity and aggregation of the mutant protein. Mutation of discreet functional domains of the AR and sites of posttranslational modification enable the detailed analysis of the role of AR function and metabolism in toxicity and aggregation of polyglutamine-expanded AR. This analysis could potentially lead to the development of targeted therapy for the treatment of SBMA. We have developed a stably transfected, tetracycline-inducible, cell model that replicates many of the hallmarks of disease, including ligand-dependent aggregation and toxicity, and provides a relatively quick system for the reliable expression and analysis of the mutated polyglutamine-expanded AR. Multiple cell lines, each expressing the androgen receptor with a distinct functional mutation, can be created and the dose of tetracycline modulated to produce equal protein expression across lines in order to evaluate the structural and functional requirements of AR toxicity and aggregation. Results from these studies can then be validated in a disease-relevant cell type, spinal motor neurons, using viral delivery of the gene of interest into dissociated spinal cord cultures. Utilization of these cell models provides a relatively rapid, cost-effective experimental pathway to analyze the role of distinct steps in AR metabolism in disease pathogenesis using in vitro systems.

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Source
http://dx.doi.org/10.1007/978-1-62703-438-8_18DOI Listing

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