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

Bonner, Eric Raymond

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Description

Title

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

Author(s)

Bonner, Eric Raymond

Issue Date

2001

Doctoral Committee Chair(s)

Switzer, Robert L.

Department of Study

Biochemistry

Discipline

Biochemistry

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

Biology, Molecular

Language

eng

Abstract

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.

Graduation Semester

2001

Type of Resource

text

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http://hdl.handle.net/2142/84775

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