Characterization of the Interactions Between Model RNA, Protein, and Bio-relevant Nanoparticles

Undergraduate #275
Discipline: Nanoscience
Subcategory: Nanoscience

Kate Inchun - LSU Shreveport
Co-Author(s): Chandler Curtis and Spencer Thomas, Missouri State University, Springfield, MO Shanna Marroquin, Miranda Hurst, and Robert Delong, Kansas State University, Manhattan, KS

RNA:Protein nanostructures such as ribosomes, spliceosomes, etc. are important for controlling gene expression. Zinc and magnesium are well known for mediating protein and RNA interactions and stability in cells and tissues. Therefore, with increased use of zinc, magnesium and other bio-element or trace element (boron) nanoparticles, the effect these have on proteins and RNA is one of the most important questions in bionanoscience today. My goal was to investigate if Protamine protein binds Poly IC RNA and if we could demonstrate an interaction between the Poly IC:Protamine with these nanoparticles that are of biological interest. Within cells, RNA and proteins interact to form nanostructures such as ribosomes, spliceosomes, etc. Nanoparticles enter cells and the effect that they have on these RNA:Protein structures is currently unknown. We have used a model RNA and protein to investigate the effects of the addition of nanoparticles. Poly IC is a double stranded RNA whose biological effects mimics viruses and possesses anti cancer properties. Protamine is a protein similar to histone proteins which bind to DNA in chromosomes, which our group has previously shown also binds RNA to form Protamine:RNA nanocomplexes. Poly IC:Protamine interaction was detected using gel electrophoresis and zeta potential.

Based on previous research from our lab and others, we selected zinc oxide (ZnO), boron carbide (B4C), and magnesium oxide (MgO) nanoparticles for their biological implications. The mean zeta potential values for these nanoparticles alone in ddH2O are: 30.1 for ZnO, -43.3 for B4C, and 9.53 for MgO. The mean values after the addition of Poly IC:Protamine are: 23.6 for ZnO, 26.9 for B4C, and 13.9 for MgO. The direction of bands found in the gel was consistent with the charges in the zeta potential readings. The fluorescence spectral signatures found also showed proof of interaction through changes in emission intensity at a given wavelength. In future research, these complexes we created would be sized and tested for which ratio of materials maintains nanosize stability.

Funder Acknowledgement(s): Funding was provided by, LSAMP National Science Foundation Grant #1305059.

Faculty Advisor: Robert Delong,