Investigation of the Molecular Interaction Between Pyr mRNA and the Bacillus Subtilis Attenuation Regulatory Protein, PyrR

Gel mobility shift experiments using progressively shorter variants of BL2 mRNA determined that the minimal RNA necessary for tight PyrR binding was 28 nt long. The stoichiometry of the PyrR-pyr mRNA interaction was determined to be equimolar using a gel mobility shift titration assay. The effects of 31 structural variants of BL2 on PyrR binding were studied. Twelve of the variations had little effect, three caused a moderate defect in binding, and sixteen severely disrupted binding. All PyrR-binding mRNAs share a conserved secondary structure, consisting of a lower stem, purine-rich internal bulge, upper stem, and terminal hexaloop, as well as two conserved sequence motifs. Variants that significantly altered the secondary structure of the mRNA disrupted binding. Variants that disrupted conserved sequences while leaving RNA secondary structure intact were tolerated in the upper stem, but disrupted binding in all other areas.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Bonner, Eric Raymond

Contributors dc:contributor

• Switzer, Robert L.

• Biology, Molecular

Language dc:language

eng

Identifier

(MiAaPQ)AAI3023022

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/84775