Mutagenic Analysis of the Expression Platform of a Model Purine Riboswitch

Juels, Parker

Undergraduate Honors Thesis

Mutagenic Analysis of the Expression Platform of a Model Purine Riboswitch Public Deposited

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Citeable URL: https://scholar.colorado.edu/concern/undergraduate_honors_theses/dn39x297f

Abstract

Riboswitches are RNA based transcription and translation controllers. What makes them unique is their ability to recognize and up or down regulate transcription or translation based upon the presence or absence of a small molecule (ligand)¹. Riboswitches have two key domains, an aptamer domain and an expression platform. The aptamer domain is responsible for binding and recognizing the ligand. The expression platform changes confirmation upon ligand binding to adapt a helix that could either halt or allow transcription and translation to occur². The PbuE adenine-responsive riboswitch isolated from Bacillus subtilis has served as a model system for understanding and studying riboswitch structure and function. The structure and function of the aptamer domain of this switch has been well established³. However, the expression is less well established. The general sequence and structure is known but its function and sequence elements need to be studied in greater detail. To determine the role of strand exchange and the importance of key sequence elements to strand exchange, mutagenic analysis of a key sequence element involved in strange exchange was conducted. The results of this analysis suggest the importance of base pairing, especially towards the end of the helix formed during strand exchange. Additionally, a sequence element connecting the aptamer domain and expression platform was studied. However, this screening did not provide usable conclusions and further screening and analysis is necessary.

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