Celebration of Scholarship and Creative Activity 2021
“An Exploration of the Relationship Between the Small Protein MntS and the MntP Export Protein Regarding Manganese Homeostasis in Escherichia coli“
Kalista Paszczak
Senior, Chemistry
Abstract
When controlled, the transition metal manganese is crucial for cellular life. However, when present in unusually high levels, manganese can restrict the ability for cells to grow. Thus, specialized export proteins, such as MntP, help combat metal toxicity induced by manganese. Recent studies have identified another new, small protein called MntS that also plays a role in maintaining homeostasis. It’s mechanism of action, however, remains unknown. This research focuses on the importance that MntP holds, as well as the potential function of MntS. First, wildtype strains of E. coli were modified to delete the mntP gene and to replace it with a gene encoding a similarly functioning export protein, known as MneA. Two different assays were used to test the importance of MntP; the first, a metal sensitivity assay, examined whether MntP was required for the cell to survive metal toxicity. The second, a two-hybrid assay, was used to determine whether MntP was required for the interaction between MntS with itself. The first set of experiments concluded that MntP is, in fact, required by the cell to survive metal toxicity induced by manganese. In contrast, the second set of experiments alluded that MntS can bind to itself even in the absence of MntP. This works provides an insight on the relationship between MntP and MntS, in the hopes of clarifying the unique function of MntS.
Project Background
Before working on this project, I didn’t know very much about E. coli and the processes it carries out to maintain homeostasis. I knew Dr. Waters from being in her biochemistry class, and upon reading about the research she was doing, I was interested not only in learning more about her work but also in potentially working with her. Because I didn’t have much background knowledge on bacteria, when I first started learning about the project, it was a little intimidating because there was so much to know! However, she explained things very clearly and precisely, and was super eager to answer any questions that I had. Once I learned a little about the work and got to start in the lab, it was cool for me to realize that many of the techniques that were used were things I had previously learned in other classes. The first few semesters consisted of preparation work, as we were trying to create a new strain of E. coli before we could start the actual experiments. We ended up finally succeeding in creating the new strain last spring (2020) and over the summer we got to start on the actual experiments. We have been pretty successful with both of our experiments – in both cases we obtained enough information to answer both of our research questions. We’ve been working really hard this semester to fine tune our second experiment, and I’m really excited to share with everyone what we’ve accomplished so far!
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