Chemistry

Systematic Optimization of Parameters of Compucell 3D For Modeling Autophagic Bodies

Garrett Bailey Steven Backues, faculty mentor

Autophagy is the process in which cells uptake cellular material and recycle it under starvation conditions, and understanding it is important for human health. Visualizing autophagic bodies by electron microscopy is an important part of this research, but is limited because it only gives flat 2D images. This can lead to slices of bodies with misleading sizes, and even exclude whole bodies. 3D modeling bypasses this as it can visualize bodies as wholes, allowing more accurate interpretation of electron microscope images. We are optimizing the parameters of the modeling program Compucell 3D to mimic the natural configuration of autophagic bodies within the cell.

Poster / Live Session 1 / 10:00 a.m.

Investigating the Binding Interaction of UHRF1 PHD D334A and UHRF2 PHD D363N to H3K9me3

Kaitlyn Bricker Brittany Albaugh, faculty mentor

Ubiquitin-like, containing PHD and RING finger domains protein 1 and 2 (UHRF1 and UHRF2) are epigenetic histone reader proteins. D334 of UHRF1 and D363 UHRF2 within the PHD is proposed to ionically interact with R2 of H3. To test this possibility, we created UHRF1 PHD D334A and UHRF2 PHD D363N mutants by mutagenesis and analyzed their impacts on the binding of H3 peptides by fluorescence polarization. We found that changing the negatively charged amino acids with neutral amino acids ceased the ability for the reader proteins to bind, furthering our understanding of these reader proteins and their connection to cancer.

Poster / Live Session 1 / 9:40 a.m.

Measuring Atg12-5 Attachment During the Autophagic Process by Western Blotting

Ayanda Brydie Steven Backues, faculty mentor

Autophagy is a degradation process in which damaged organelles or cellular components are digested to maintain homeostasis within the cell. This project will focus on nonselective or bulk autophagy, initiated when the cell is starved of nutrients and mediated by Atg proteins. Our current work is part of the broader examination of the effect Atg10 has on autophagosome size and number. Atg10 is responsible for the attachment of Atg12 to Atg5, which is essential for the formation of autophagosomes. The attachment of Atg12 to Atg5 will be verified in Atg10 mutants by western blotting.

Poster / Live Session 1 / 10:20 a.m.

Characterization of the Histone Binding Properties of D337N UHRF1 Mutant

Nick Butkevich Brittany Albaugh, faculty mentor

UHRF1 is an epigenetic protein involved in cancer regulation. The binding of UHRF1 protein to histone H3 via ionic interactions may cause the repression of tumor suppressor genes. The goal of this project is to determine the role of amino acid D337 in UHRF1 in binding to histone H3. A D337N UHRF1 mutant (Aspartic Acid to Asparagine) was generated by PCR. The mutated DNA was then transformed and expressed in Rosetta E. Coli. The mutant protein was purified and the binding affinity of the mutated proteins to histone H3 peptide were measured by fluorescence polarization. The results of this work will be presented

Poster / Live Session 1 / 9:20 a.m.

Jessica K. Callaghan Gavin D. Edwards, faculty mentor

For the last decade the atmospheric chemistry group at EMU has measured the concentration of the important pollutant ozone. This EPA regulated gas is known to cause damage to human health such as causing breathing problems for sensitive groups. In early 2020, the Covid 19 global pandemic resulted in the chance to examine the effect of the lockdown on local and regional air quality with respect to ozone. It was hypothesized that there could be a change in ozone levels measured during lockdown relative to a 10-year measurement average. In this poster we report the findings for ozone levels in 2020, and discuss the impact of Covid 19 on our air quality.

Poster / Q&A Session 2

Mutated Autophagic Proteins and Their Binding Partners

Chase Chitwood and Sophia Taylor Steven Backues, faculty mentor

Autophagy is a cellular process in which cellular components are broken down and recycled. Autophagy is important for maintaining healthy cells and may play a role in certain illnesses and disorders. Many proteins are involved in this process and we are researching how these proteins interact. In the lab, we are using molecular cloning techniques to produce yeast with DNA that expresses mutated autophagic proteins to determine how these mutations impact protein interactions. Using a method termed co-immunoprecipitation, we are testing protein-protein interactions and determining if certain mutations cause loss of interaction between the two expressed proteins.

Poster / Live Session 1 / 9:50 a.m.

Payton Dunning Steven Backues, faculty mentor

Faced with starvation, eukaryotic cells will recycle cellular material for reuse through the process of autophagy. In yeast cells, cellular material is transported to the vacuole in autophagosomes to be degraded. The resulting autophagic bodies within the vacuole can be measured using electron microscopy to generate 2D slices or snapshots of the cell vacuole. These 2D images are not perfect representations and may leave out entire autophagic bodies. To better understand the size, number, and configuration of autophagic bodies within vacuoles undergoing autophagy, this project seeks to develop software for simulating and analyzing 3D autophagic bodies within a cell vacuole.

Poster / Live Session 1 / 9:30 a.m.

Systematic Studies Towards the Development of a Novel Beer Style – Smoked IPA

Clint Lafferty Cory Emal, faculty mentor

The explosion of the craft beer industry in recent years has led to the creation of numerous new beer styles, including variations on the India Pale Ale (IPA), a style of beer defined by a hop-forward flavor profile. The majority of modern variations of IPA focus on the characteristics of the hops themselves (e.g. pine, fruit), often paired with fruit and/or a sour component. Smoke, a historically important flavor, has been overlooked. In collaboration with a local brewery, we have developed a recipe that combines these divergent flavors. By evaluating multiple smoked malts alongside a range of hop varieties, we have developed a prototype recipe that will help establish this style of beer.

Poster / Live Session 2 / 11:15 a.m.

Isaak Miller Brittany Albaugh, faculty mentor

Epigenetics explores the changes caused by modifications of gene expression without altering the DNA sequence. UHRF2 is a protein involved in cell-cycle regulation and is thought to have possible anticancer characteristics. The first project involved creating a DNA construct that expresses UHRF2 protein in E.Coli cells. The second project involved the purification of the UHRF2 protein construct by chromatography. The goal of the project is to determine the crystal structure of UHRF2 to further our understanding of the mechanism by which UHRF2 interacts with histone H3.

Oral / Q&A Session B

Replacement of Hydrazide Moieties in Inhibitors of Plasminogen Activator Inhibitor-1

Micaela Schempf Cory Emal, faculty mentor

The human protein plasminogen activator inhibitor-1 (PAI-1) is largely responsible for the regulation of fibrinolysis, or blood clotting. Mammalian PAI-1 concentrations are typically very low, but abnormally high levels of PAI-1 correlate with a number of common pathological conditions, including stroke, heart attack, and kidney fibrosis. The ability to target and inhibit excess PAI-1 with a selective small-molecule inhibitor in order to treat or prevent these pathologies is desirable. This study focuses on the replacement of the hydrazide functional group in existing PAI-1 inhibitors with functionalities that potentially retain activity and provide improved pharmacokinetic properties.

Poster / Live Session 1 / 10:10 a.m.

The Effect of Autophagy Related Protein 10 and its Mutants on Autophagosome Size and Number

Nadia Silvia Steven Backues, faculty mentor

Autophagy is the cellular degradation process in which cellular contents are encapsulated by double-membrane vesicles, autophagosomes, and delivered to the vacuole to be degraded and recycled. This process is important for cell health and homeostasis. There are approximately 32 different autophagyrelated proteins involved. Atg10 is an enzyme that may affect overall autophagic activity by changing autophagosome size and/or number. Our goal is to find mutants that cause a partial loss in Atg10’s activity by mutating some residues near the active site and testing the functionality of these mutants by performing western blots and enzymatic assays.

Poster / Live Session 1 / 9:00 a.m.

Air Quality on the EMU Campus During the Covid 19 Lockdown

Katrina White Gavin D. Edwards, faculty mentor

The first lockdown associated with the Covid 19 pandemic has offered atmospheric chemists the opportunity to look at the changes in pollution associated with the impact of the lockdown on our local air quality. In this work, we will report the impact of the lockdown on air quality measured on the EMU campus. We show the changes in one specific EPA regulated pollutant, PM2.5. This poster will discuss the changes in the concentration of this aerosol over the lockdown period by comparing our measurements to those of previous years, and the implications for improving local and regional air quality in the future.

Poster / Q&A Session 2