Biochemistry, Biophysics, and Structural Biology Commons^™

Tail-Tape-Fused Virion And Non-Virion Rna Polymerases Of A Thermophilic Virus With An Extremely Long Tail, Anastasiia Chaban, Leonid Minakhin, Ekaterina Goldobina, Brain Bae, Yue Hao, Sergei Borukhov, Leena Putzeys, Maarten Boon, Florian Kabinger, Rob Lavigne, Kira Makarova, Eugene V Koonin, Satish Nair, Shunsuke Tagami, Konstantin Severinov, Maria Sokolova

Department of Biochemistry and Molecular Biology Faculty Papers

Thermus thermophilus bacteriophage P23-45 encodes a giant 5,002-residue tail tape measure protein (TMP) that defines the length of its extraordinarily long tail. Here, we show that the N-terminal portion of P23-45 TMP is an unusual RNA polymerase (RNAP) homologous to cellular RNAPs. The TMP-fused virion RNAP transcribes pre-early phage genes, including a gene that encodes another, non-virion RNAP, that transcribes early and some middle phage genes. We report the crystal structures of both P23-45 RNAPs. The non-virion RNAP has a crab-claw-like architecture. By contrast, the virion RNAP adopts a unique flat structure without a clamp. Structure and sequence comparisons of …

Fused In Sarcoma Regulates Glutamate Signaling And Oxidative Stress Response, Chiong-Hee Wong, Abu Rahat, Howard C Chang

Rowan-Virtua School of Osteopathic Medicine Departmental Research

Mutations in fused in sarcoma (fust-1) are linked to ALS. However, how these ALS causative mutations alter physiological processes and lead to the onset of ALS remains largely unknown. By obtaining humanized fust-1 ALS mutations via CRISPR-CAS9, we generated a C. elegans ALS model. Homozygous fust-1 ALS mutant and fust-1 deletion animals are viable in C. elegans. This allows us to better characterize the molecular mechanisms of fust-1-dependent responses. We found FUST-1 plays a role in regulating superoxide dismutase, glutamate signaling, and oxidative stress. FUST-1 suppresses SOD-1 and VGLUT/EAT-4 in the nervous system. FUST-1 also regulates synaptic AMPA-type glutamate receptor …

Lipid Genetics In Pregnant Women: Maneuvering The Challenges Of Using Degraded Samples, Kylee Meece

Mahurin Honors College Capstone Experience/Thesis Projects

The results of this project will explore a novel approach to lipid metabolism in pregnant women by exploring the relationship between maternal gene expression of genes critical for fat metabolism, protein expression, and maternal and neonatal outcomes. The study will also examine gene expression in lean pregnant women compared to obese pregnant women, shedding light into the adverse pathology of obesity during pregnancy. These findings may uncover a mechanism related to gene-expression that connects unfavorable maternal fat metabolism to inflammation and/or insulin resistance, which may help to uncover future targets for interventions.

The analysis of lipid gene expression requires the …

Thymidine Phosphorylase Is A Promising Target In Sars-Cov-2 Spike Protein-Enhanced Thrombosis, Renat Roytenberg

Theses, Dissertations and Capstones

Thrombosis, as an underlying mechanism in cardiovascular diseases, is the leading global cause of death in the 21st century. Infection is a risk factor for thrombosis, and the coronavirus disease 2019 (COVID-19) pandemic has led to a rise in the incidence of thrombosis, thereby increasing mortality. In comparison to previous severe coronavirus outbreaks, rates of thrombosis in COVID-19 have been unprecedented. COVID-19 is caused by the novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 spike protein (SP), the key used to enter host cells, is implicated in thrombosis pathology, though mechanisms are not fully understood. The SARS-CoV-2 …

Assessing The Role Of Kmt2b Knockdown In Rhabdoid Tumorigenesis Utilizing Crispri, Karah Edmonds, Andrea Florian Phd

Science University Research Symposium (SURS)

An aggressive blood cancer commonly observed in pediatric patients, Mixed Lineage Leukemia (MLL), has a regulatory MLL complex that promotes histone methyltransferation. Modifying histones utilizing histone methyltransferases like KMT2B (MLL2), is essential for gene expression by regulating transcription. Loss of function and inhibition of KMT2B leads to dysregulation of transcription and therefore promotes MLL tumorigenesis. To improve cancer-targeted therapies, we will use CRISPR interference (CRISPRi) to knock down the expression of KMT2B and measure levels of Rhabdoid tumor cell proliferation and apoptosis. Previous research surrounding the MLL complex suggests that decreased expression of KMT2B will lead to increased levels of …

Analysis Of Protein-Protein Interactions Between Legionella Pneumophilia Effectors, Briana Harness

Mahurin Honors College Capstone Experience/Thesis Projects

Legionella pneumophila an intracellular, opportunistic pathogen that is the causative agent of Legionnaires’ disease and Pontiac fever. A key component of Legionella’s pathogenicity is a series of effector proteins, which aid in the establishment of the Legionella-containing vacuole (LCV). The abundant library of effector proteins L. pneumophila has within its arsenal gives L. pneumophila the ability to infect several different hosts. The pathogenesis of L. pneumophila within various hosts depends on its ability to modulate the activity of its effector proteins through protein-protein interactions within effectors and eukaryotic host cell proteins. By understanding how L. pneumophila regulates its effectors, a …

Detecting Interactions Between Mps3 In Linc Complex And Ndj1 In Nuclear Pore Complex In Mechanically Stressed Yeast, Dean Boecher, Rebecca Adams

[Archive] Belmont University Research Symposium (BURS)

The mechanisms of how mechanical stress is translated into cellular action and structural reorganization within the nuclear envelope are largely unexplored. The Linker of Nucleoskeleton and Cytoskeleton (LINC) complex is a transmembrane protein complex that connects the actin cytoskeleton to the lamin nucleoskeleton, enabling mechanical forces to be translated between the cytoplasm and the nucleus. A better understanding of the translation of physical forces into cell responses can be gained through confirming the existence of interactions between LINC complex proteins and nuclear pore complexes (NPCs), which control the exchange of biochemical signals and macromolecules in and out of the nucleus …

Long-Read Mdm4 Isoform Sequencing Reveals Aberrant Isoform Landscape In Metastatic Melanomas, Nehaal Rooha Patrick

Browse all Theses and Dissertations

The changes in splicing of MDM4 in human melanomagenesis are critical to p53 activity and represent potential therapeutic targets. Genomic characteristics of melanoma and precursor lesions, such as copy number variation and gene expression, have been investigated in detail. However, there has never been a detailed survey of splicing changes that occur during melanomagenesis. Splicing changes can be a very early event in melanoma tumor progression, with characteristic changes in the p53 pathway already in place in early nevi. MDM4 is upregulated in a strong majority of melanoma cases and has been described as a “key therapeutic target in cutaneous …

Role Of Insulin-Like Growth Factor-1 Signaling In Diet-Enhanced Bone Elongation, Cassaundra A. White

Theses, Dissertations and Capstones

Childhood obesity has increased precipitously to the point that almost 20% of children between the ages of 2 and 19 have been diagnosed with obesity. While obesity is a complex metabolic disorder, one contributing factor is diet, particularly a diet high in fat. Among many problems associated with diet-induced obesity, the impact on skeletal health is often overlooked. In addition to increased stress on the skeleton due to excess weight, obesity also disrupts the endocrine and immune systems that, in turn, can secondarily alter skeletal growth. Obesity during the early years of childhood is particularly detrimental because the skeleton is …

Microbial Prevalence And Antibiotic Resistance Associated With Urban Food Production Systems, Nirosha Ruwani Amarasekara

Wayne State University Dissertations

Despite the increasing popularity of urban agriculture and farmers’ markets, there is a lack of scientific data on microbial prevalence and antibiotic resistance associated with this unique food production system. The overall goal of this research is to understand antibiotic resistance and microbial prevalence in soil and fresh produce associated with urban agriculture. Specifically, we aim to investigate ARG in association with resistance genes to metals and other compounds in urban agricultural soil as well as antibiotic resistome and Listeria contamination in fresh produce distributed in urban communities.

In the initial phase of this research, soil samples were collected from …

The Role Of Ferroptosis In Cholangiocarcinoma, Apiwit Sae-Fung

Chulalongkorn University Theses and Dissertations (Chula ETD)

Cholangiocarcinoma (CCA) is a highly heterogeneous and aggressive malignancy of the bile ducts, characterized by a poor prognosis and high mortality rate. Despite advancements in treatment, effective targeted therapies and reliable prognostic biomarkers remain limited. Ferroptosis, a regulated form of cell death, has emerged as a potential therapeutic target in various cancers. However, its role in CCA remains poorly understood. In this study, we conducted a comprehensive analysis of ferroptosis-related genes (FRGs) to predict the prognosis of CCA and identify potential therapeutic targets. Using the least absolute shrinkage and selection operator (LASSO) Cox regression analysis on a training dataset, followed …

The Role Of Uchl1 In Skeletal Muscle Development And Regeneration, Ryan Antony

Dissertations and Theses

Ubiquitin C-terminal hydrolase L1 (UCHL1) is a deubiquitinating enzyme that was originally discovered in neurons. UCHL1 is also expressed in skeletal muscle, but its functions remain to be fully understood. Myogenesis is a critical process involved in embryonic development, growth, and regeneration following injury. Skeletal muscle injury is prevalent in trauma and surgical procedures, and skeletal muscle ischemia-reperfusion (IR) injury is a common yet dangerous public health problem. Here we reported that UCHL1 negatively affects muscle growth during aging as well as the regeneration process following IR injury. First, we observed that UCHL1 knockdown in C2C12 myoblasts resulted in increased …

Mathematical Modeling Of Microscale Biology In Polyelectrolyte Brushes, William J. Ceely

CGU Theses & Dissertations

Biological macromolecules including nucleic acids, proteins, and glycosaminoglycans are typically anionic and can span domains of up to hundreds of nanometers and even micron length scales. The structures exist in crowded environments that are dominated by multivalent electrostatic interactions that can be modeled using mean-field continuum approaches that represent underlying molecular nanoscale biophysics. In this thesis, we develop such models for polyelectrolyte brushes using both steady state modified Poisson-Boltzmann models and transient modified Poisson-Nernst-Planck models that incorporate important ion-specific (Hofmeister) effects. The transient model enables observation of the relative physical effects as an initial non-equilibrium state relaxes to the steady …

Climate Change And Coastal Development Impacts On Oyster Abundances In Mosquito Lagoon, Fl, Emily Suchonic

Graduate Thesis and Dissertation 2023-2024

Live eastern oyster (Crassostrea virginica) reefs have declined by 62.6% in Mosquito Lagoon (ML) along the eastern Florida coast since 1943. While this species creates reefs by successive generations of oysters recruiting to conspecific shells, C. virginica can also attach to non-reef substrates including mangrove roots and armoring (e.g., seawalls), which may help counteract reef habitat loss. In recent decades, warmer winters have enabled red (Rhizophora mangle) and black (Avicennia germinans) mangrove expansion in subtropical salt marshes and temperate estuaries where oyster reefs occur. Additionally, 11.8% of ML's shorelines have been armored as of …

Uracil Landscape Of Mouse B Cell Genome Undergoing Class-Switch Recombination, Fathima Rukshana Mohamad Ramshan

Wayne State University Dissertations

Activation-induced cytosine deaminase (AID) is required for both somatic hypermutation (SHM) and class switch recombination (CSR) within mammalian antibody genes. Uracils created by AID in the Ig loci are recognized and processed by base-excision and mismatch repair pathways leading to antibody diversification. This involves a hypermutation process restricted to the V(D)J segments of the immunoglobulin genes, SHM, and an error-prone end-joining process that causes isotype switching, CSR, but the molecular details of these processes have remained obscure for the past 20+ years. Most studies of SHM and CSR have used AID-promoted mutations as a proxy for the uracils, but their …

A New Model For Polar Growth In Mycobacteria: The Role Of Wag31 And Mmpl3 In Cell Wall Synthesis, Neda Habibi Arejan

Biology Dissertations - Archive

The disease Tuberculosis (TB) is caused by the pathogen Mycobacterium tuberculosis and was the cause of over one million deaths worldwide in 2023 (World Health Organization, 2023). TB treatment is difficult due to antibiotic tolerance and emerging resistance in Mtb strains. Mycobacterial cells have different regulatory mechanisms that allow them to adjust to myriad environmental stressors: these responses often lead to antibiotic tolerance. Therefore, studying responses to stress will lead to insights about antibiotic tolerance and treatment failure. Here, we specifically study two essential mycobacterial proteins, Wag31 and MmpL3, involved in cell wall metabolism. We show that Wag31 has distinct …

Osteohistological Assessment Of Edmontosaurus Radii From The Liscomb Bonebed, Prince Creek Formation, North Slope, Alaska, Emma Griffin

Student Research Poster Presentations 2024

Hadrosaurs were one of the most diverse dinosaur groups in the Late Cretaceous period and are known for their numerous articulated skeletons and monodominant bonebeds (Wosik et al., 2020). They are more commonly known as duck-billed dinosaurs and are found globally including latitudinal extremes such as the artic and Antarctica. Edmontosaurus, a genus of hadrosaur, are herbivores and are very large growing up to 8,000 pounds. Edmontosaurus were known to travel in herds as a defense mechanism and were one of the last non-avian dinosaurs living during the late Campanian stage through the end of the Maastrichtian stage, right up …

Rhizophora Mangle (Red Mangrove) Seedling Success In Different Habitats In Mosquito Lagoon, Florida, Usa, Mekail N. Negash

Graduate Thesis and Dissertation 2023-2024

Mangroves provide many ecosystem services in coastal environments around the world. These include water quality improvement, creating habitats for terrestrial and aquatic species, and stabilizing shorelines. In central Florida, the red mangrove Rhizophora mangle is a common species in coastal wetlands, and recently the number of individuals successfully recruiting to intertidal oyster reefs has greatly increased, possibly because biogeochemical hot spots are present on oyster reefs due to nutrient-rich biodeposits from the live oysters. To understand how well R. mangle responds in terms of survival and growth to the suite of variables associated within these two unique habitats, I tracked …

Synthesis Of Dihydro-Β-Carbolines By Tandem C-H Activation And Aza-Michael Reaction In The Presence Of Rh(Ii)/Squaramide Catalysis: Investigation Of The Mechanism Via Computational Methods, Syed Wasiuddin

Senior Honors Theses and Projects

N-heterocycles are the major component of various bioactive natural products, pharmaceuticals, and their precursors. In particularly, β-carbolines are examples of substituted indole and its derivatives that serve as the basic structures of diverse, therapeutically important molecules with broad pharmacological properties. Rajasekar and Anbarasan have suggested a novel one-pot synthesis for dihydro-β-carbolines involving more than one catalytic system: a rhodium acetate dimer for C-H insertion on the indole and a basic organo-catalyst for the subsequent aza-Michael addition. The product can be obtained enantioselectively by using a chiral squaramide as the organo-catalyst. Since the mechanism of this reaction, including the rate determining …

Feeding Systems And Fresh Lamb Color Stability, Koushik Mondal

Theses and Dissertations--Animal and Food Sciences

Consumers consider the surface color of fresh lamb as the indicator of freshness and wholesomeness, which affects their purchase decisions. The redox chemistry of myoglobin (Mb) determines the color of fresh lamb. Artificial raising on milk replacer (a pre-weaning strategy) as well as red clover supplementation are management strategies used in lamb production. This thesis focuses on the effects of artificial raising on milk replacer and red clover supplementation on the color and oxidative stability of fresh lamb.

In the first experiment, the effect of pre-weaning with milk replacer on the carcass attributes, color stability and lipid oxidation of lamb …