Using fluorescent timer to study protein inclusion biogenesis and removal by autophagy.

• Main Author: Lee, Pei Yu.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2012
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/49399
• Institution: Nanyang Technological University

Protein inclusion, formed from the aggregation of misfolded proteins can be degraded by autophagy. Autophagy has been shown to be a selective process dependent on the nature of the protein inclusions. Research on whether and how the aggregation pattern of an aggregate-prone protein influences its handling in autophagy has not been extensively researched on. This project aims to use autophagy-amenable A38 protein and autophagy-resistant p38 protein fused with fluorescent timer to observe spatial and temporal behaviour of the newly synthesized proteins and the older proteins, and understand their aggregation and clearance patterns. We found that aggregates and aggresomes of A38-pmed-FT are indeed cleared by induced autophagy while p38-pmed-FT aggregates and aggresomes remained uncleared or minimally cleared. We also found that while aggregates of both FT fused proteins have no distinct distribution of the newer and older proteins, the aggresomes of these proteins showed some difference. The newer proteins of p38-pmed-FT are preferentially localized in the center of its aggresome while the distributions of older and newer proteins in aggresome of A38-pmed-FT are more evenly spread. This result indicate some difference in how the aggregates of the different proteins come together to form aggresome which might affect its clearance susceptibility.