Utilization of plant Puf RNA-binding proteins to manipulate endogenous mRNA physiology
Date
2019-02-01
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Abstract
RNA-binding proteins have an important role in the regulation of post-transcriptional gene expression. The Puf (Pumilio) family of RNA-binding proteins bind to RNAs in a sequence specific manner. The RNA-binding domain of canonical Puf proteins (the Pumilio homology domain, PUM-HD) consists of eight Puf repeats. Each Puf repeat binds to RNA in a modular fashion, so that one Puf repeat binds to a single nucleotide. The nucleotide binding recognition code of Puf repeats has been well characterized. This one repeat:one nucleotide recognition code has allowed for the alteration of Puf repeats so that the PUM-HD can bind specific RNA targets. These altered Puf repeats have been fused to effector domains so that fusion proteins can affect RNA physiology in the cell. The aim of this thesis was to fuse an Arabidopsis Puf RNA-binding protein with unique binding characteristics to two types of effector domains. These fusion proteins were expressed in onion epidermal cells to determine if they could alter the stability or translation of a reporter mRNA. This research provides foundational evidence that highlights the potential for the utility of plant Puf proteins to alter gene expression for applied purposes
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Keywords
APUM23, Puf proteins, RNA-binding proteins, Protein engineering
Citation
Wen, X. (2019). Utilization of plant Puf RNA-binding proteins to manipulate endogenous mRNA physiology (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca.