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       In my research laboratory I have two broad areas of interest:

 

      The first is focused on the synthesis of proteins. Protein synthesis is an absolute requirement for growth and development in biological organisms, yet our understanding the core component (the ribosome) and its individual constituents (ribosomal proteins) in plants remains limited. However, we have been able to demonstrate that there are some distinct differences between a few r-proteins found in plants, but not in other organisms. The 12-kDa acidic ribosomal phosphoproteins (P-proteins) form a universally conserved structure in the active site of the large ribosomal subunit (60S) and are thought to assist in multiple phases of protein synthesis. Higher plant ribosomes possess evolutionarily conserved (P1, P2a and P2b) and evolutionarily distinct plant specific (P3) forms of 12-kDa P-proteins. The presence, abundance and phosphorylation of the 12-kDa P-proteins of maize ribosomes varies during development. Therefore, I hypothesize that the heterogeneity observed with these proteins plays a role in the regulation of gene expression via selective mRNA translation. The objectives of this research are to determine the organization of acidic ribosomal P-protein stalk and to elucidate the functional significance of ribosome heterogeneity with respect to the acidic ribosomal P-proteins in the model plant system, Arabidopsis thaliana. We have been utilizing a technique which will allows us to sort ribosomes based on their protein composition. Once this sorting has been accomplished we can extract mRNAs from the sorted ribosomes and sequence the mRNAs to determine if ribosomes with varying P-protein stalk composition, translate a different set of mRNAs. This technique will also be applied to another ribosomal protein (S15a Type II), which also appears to be plant specific. Together, these data will provide a landmark for the analysis of plant ribosomes and has the potential to widely impact agriculture through the prospect of higher crop yield.

 

The second area of my research stems from a collaboration with Dr. Antje Lauer at CSUB, in which we hypothesize that we will find powerful antifungal metabolites produced by cutaneous bacteria of amphibians given that cutaneous bacteria of amphibians are able to protect their host against pathogens (including the recently emerged fungal pathogen Batrachochytrium dendrobatidis) and that antibiotics from naturally occurring compounds have been used to combat infectious diseases. This project focuses on several different aims. 1) To characterize and compare cutaneous bacterial diversity of R. catesbeiana (North American Bullfrog) and B. b. halophilus (Boreal Toad) by analyzing amplified 16S rDNA fragments from DNA isolated from amphibian skin swabs using Denaturant Gradient Gel Electrophoresis (DGGE). 2) To isolate members of the skin microbiota and challenge them against different environmental fungi. 3) To identify the antifungal bacterial species by sequencing a portion of the 16S rRNA gene. The data we generate has the potential to widely impact human health issues by identifying potential antibiotics or novel chemotherapeutic compounds.

 

kszick-miranda@csub.edu