Understanding the RNA-Protein Contacts Required for Successful Infection of the ssRNA Bacteriophage Qβ
Abstract
The single-stranded (ss) RNA bacteriophage Qβ, which infects piliated bacteria, most commonly Escherichia coli, requires several RNA-protein interactions for successful infection. Based on modeling the RNA secondary structure of the genome into the electron density of cryo- EM structures, interactions thought to be critical for infection were seen between RNA stem- loops at the 3’ end of the genome (U1 and R1 stem loops) and the maturation protein. The purpose of this study was to validate these proposed interactions. Mutations were introduced into the R1 and U1 stem loops that change the quality of the interactions with the maturation protein, and the resulting virus-like particles’ infectivity was observed. The importance of RNA-protein interactions will be further explored by swapping the maturation protein of the closely related bacteriophage MS2 with that of Qβ. If genome-protein interactions similar to those seen in infectious Qβ are conserved, we predict that infectious virus- like particles will be produced. The goal of this project is to understand the RNA-protein interactions required by Qβ to establish infection in order to exploit these to use Qβ as a means of RNA delivery into bacterial cells. The ability to deliver engineered RNA molecules into bacterial cells has important therapeutic and industrial applications. RNA encoding bactericidal proteins or short interfering RNA (siRNA) fragments that interfere with critical gene expression can be used to suppress bacterial growth, which holds promise as a method of treating bacterial infections. Bacterial cell machinery can also be harnessed to produce proteins of interest by delivering RNA that encodes for them.
Citation
Kakwan, Haania (2020). Understanding the RNA-Protein Contacts Required for Successful Infection of the ssRNA Bacteriophage Qβ. Undergraduate Research Scholars Program. Available electronically from https : / /hdl .handle .net /1969 .1 /196679.