As I mentioned before, I am currently working in the Brennan Lab, a Biochemistry lab that has previously done extensive research into the trehalose pathway within fungi. In order to lead into my project, it is important to understand more about certain research in the Brennan lab and what exact implications it has in the large scheme of things.
Throughout recent times, immunocompromised host populations, whether transplant recipients, cancer victims, and other ICU patients (Intensive Care Unit), have continuously been invaded and killed by fungal diseases. In a search to identify antifungal targets, the Brennan lab investigated the trehalose pathway due to its fungicidal characteristics, broad relevance to many different types of fungi, and its low toxic consequences in mammalian species.
Looking a bit closer into this research, the trehalose pathway in fungi is regulated by two important enzymes that basically form the backbone of trehalose formation: Tps1 (trehalose-6-phosphate synthase) and Tps2 (trehalose-6-phosphate phosphatase). Tps1 creates trehalose-6-phosphate (t6p) from glucose-6-phosphate while Tps2 ultimately forms trehalose from t6p. Trehalose is extremely important in fungi survival as it has implications to energy reserves during times of stress and is crucial to carbon metabolism and regulation. Tps2 has been identified, due to structural research and animal tests, to be an ideal fungicidal target as mutant Tps2 fungi cannot survive in high stress environments. However, recent research has also uncovered that Tps1 and Tps2 seem to serve other functions within fungi than simply the trehalose path way, signaling that these are possible “moonlighting” proteins.
Well, you’re probably wondering, how does all of this relate to plants and my project? Recently, collaborators of the Brennan lab have studied a protein in maize, RAMOSA3 (RA3), that is part of the trehalose pathway in plants. RA3 serves the purpose of converting trehalose-6-phosphate into trehalose, an analogous function to that of Tps2 in fungi. Yet, when RA3 was mutated in maize, the phenotype of the maize changed and at the stem of the maize, and weird abnormally long branches would begin to form. This gives some indication that RA3 is much more than just a regulator within the trehalose pathway. Once again, this RA3 enzyme is suspected to have possible transcription regulation implications. My project is focused on taking the plasmid containing the RA3 protein and materials sent by collaborators to express the protein, purify it, confirm its function, and ultimately create a structure of it. In order to better understand the functions of RA3 and its implication or analogous functions to the possible fungi trehalose path, we need a literal structure of the enzyme to study substrates and other similar patterns. The moonlighting similarity is yet another thing to be studied. As RA3 is relatively still not understood, my project is focused on create a better familiarity with this enzyme.
Till next time,
Luke Sang
