Life Sciences Commons^™

Characterizing A Novel Cocksackievirus B3 Protease Mutant And Its Response To Polyamine Depletion, Bridget Hulsebosch

Master's Theses

Enteroviruses, including Coxsackievirus B3 (CVB3), are pervasive pathogens that cause significant disease, including cardiomyopathies. Unfortunately, no treatments or vaccines are available for infected individuals. We identified the host polyamine pathway as a potential drug target, as inhibiting polyamine biosynthesis significantly reduces enterovirus replication in vitro and in vivo. Here, we show that CVB3 is sensitive to polyamine depletion through the polyamine analog diethylnorspermidine (DENSpm) which enhances polyamine catabolism through induction of polyamine acetylation. We demonstrate that CVB3 acquires resistance to DENSpm via mutation of the 2A protease, which enhances proteolytic activity in the presence of DENSpm. Resistance to DENSpm occurred …

Assessing The Role Of The Interaction Between Polyamines And Heparan Sulfate In Coxsackievirus B3 Viral Infection, Oreoluwa Omoba

Master's Theses

Non-polioenteroviruses are highly infectious viruses that typically cause mild asymptomatic cases but can cause severe disease such as aseptic meningitis and dilated cardiomyopathy. Coxsackievirus B3 (CVB3) is a member of this group and is prevalent in the United States and Eastern Asia. Viruses like CVB3 rely on the host cell for many molecules in order to replicate. One such set of molecules is the polyamines. Polyamines are small, positively charged molecules that play a role in a multitude of cellular processes including RNA/DNA stabilization, gene expression, translation, and regulating membrane fluidity. Previous studies have shown CVB3 relies on polyamines for …

Understanding The Role Of Polyamine N-Acetyltransferase In The Fission Yeast Response To Perturbation Of The Cytokinetic Machinery, Mythili Ravishankar

Electronic Thesis and Dissertation Repository

The mechanisms that ensure cytokinetic fidelity are important for cellular proliferation and the maintenance of genomic integrity. To better understand these mechanisms, the actin depolymerizing drug, LatA, was previously used to screen for fission yeast genes required to prevent cytokinesis failure. This screen identified the pna1 polyamine-N-acetyltransferase. Here, I show that pna1Δ mutants display severe growth defects when exposed to even low doses of LatA. Furthermore, LatA-treated pna1Δ cells display increased rates of cytokinesis failure characterized by fragmentation of the cytokinetic actomyosin ring. Surprisingly, Pna1-GFP fusion proteins form cytoplasmic filaments that may represent cytoophidia (meaning "cellular snakes"), a class of …

Quantification Of Mitochondrial Zinc Homeostasis And Analysis Of Zinc And Polyamine-Mediated Axonal Trafficking, Dylan Hunt Fudge

Electronic Theses and Dissertations

The zinc ion is an important emerging signaling molecule for biological processes. In this work we engineered improved zinc sensors based on our previously developed fluorescent sensor GZnP1 to provide sensors with a higher fluorescent readout, faster kinetics, and a superior sensitivity to zinc. We utilized these zinc sensors and further developed the sensors to answer questions pertaining to biological zinc. We showed the labile zinc concentration in the mitochondrial matrix was less than 1 pM while the labile zinc concentration in the cytosol and mitochondrial IMS were comparable (~100 pM) across four different cell lines. Using these zinc sensors …

Integrated Molecular Characterization Of Lung Adenocarcinoma With Implications For Immunotherapy, Nicholas T. Gimbrone

USF Tampa Graduate Theses and Dissertations

This dissertation covers a variety of the genetic and molecular abnormalities of lung adenocarcinoma with an emphasis on STK11 loss and its implications on immunotherapy response. Given that lung cancer is the leading cancer killer, novel therapies are in great demand. In particular, immunotherapy has shown some of the most promise in the last decade but remains limited due to nearly 80% of patients not significantly responding. This dissertation aims to molecularly characterize lung adenocarcinoma while attempting to explain the reason why patients with STK11 loss do not respond to immunotherapy.

In the first chapter we discuss the relationship between …

Understanding Immunometabolic And Suppressive Factors That Impact Cancer Development, Rebecca Swearingen Hesterberg

USF Tampa Graduate Theses and Dissertations

The immune system is responsible for surveillance against pathogens and malignancies and adoptive transfer of antigen-specific T is a powerful new cancer treatment option. There is a clinical need to understand more about the normal regulatory processes that prevent immunosurveillance and how to manipulate this for clinical benefit. This project aims to understand T cells from three different angles. First, the biological role of the negative regulator cereblon (CRBN) was investigated in CD8+ T cells and my studies showed that higher levels of specific derivatives downstream of the amino acid glutamine may be beneficial to T cells in the immune …

Modulation Of Coxsackievirus Protease Activity By Polyamines, Courtney Noelle Dial

Master's Theses

Coxsackievirus type B (CVB3) is one of the six serotypes of the Coxsackievirus family of non-enveloped, linear, and positive-sense single-stranded RNA viruses. It is a pathogenic enterovirus that belongs to the same genus as the notable pathogen poliovirus. CVB3 can cause a range of illnesses from a fever to gastrointestinal distress but is most noteworthy for the ability to cause viral myocarditis, a swelling of the heart muscle. Coxsackievirus, like all RNA viruses, tends to develop mutations rapidly due to its error prone polymerase and lack of proofreading activity. These mutations can be advantageous for the virus, allowing it to …

The Role Of Polyamines In Enteroviral Attachment, Thomas Mckenzie Kicmal

Master's Theses

Polyamines are small polycationic molecules with flexible carbon chains that are found in all eukaryotic cells. Polyamines are involved in the regulation of many host processes and have been shown to be implicated in viral replication. Depletion of polyamine pools in cells with FDA approved drugs restricts replication of diverse RNA viruses. Viruses can exploit host polyamines to facilitate packaging, transcription, translation, and protease activity but other mechanisms remain largely unknown. Picornaviruses, including Coxsackievirus B3 (CVB3), are sensitive to depletion of polyamines and remain a significant public health threat. We employed CVB3 as a model system to investigate a potential …

Studies Of Norspermidine Uptake In Drosophila Suggest The Existence Of Multiple Polyamine Transport Pathways, Michael Dieffenbach

Honors Undergraduate Theses

Polyamines are a class of essential nutrients involved in many basic cellular processes such as gene expression, cell proliferation, and apoptosis. Without polyamines, cell growth is delayed or halted. Cancerous cells require an abundance of polyamines through a combination of synthesis and transport from the extracellular environment. An FDA-approved drug, D,L-α-difluoromethylornithine (DFMO), blocks polyamine synthesis but is ineffective at inhibiting cell growth due to polyamine transport. Thus, there is a need to develop drugs that inhibit polyamine transport to use in combination with DFMO. Surprisingly, little is known about the polyamine transport system in humans and other eukaryotes. Understanding the …

An Rnai Screen To Identify Components Of A Polyamine Transport System, Adam J. Foley

Honors Undergraduate Theses

Polyamines, specifically putrescine, spermidine, and spermine, are small cationic molecules found in all organisms. Cells can biosynthetically make these molecules, or alternatively, they can be transported from the extracellular environment. Malignant cells have been shown to require relatively high amounts of polyamines. There is a chemotherapeutic agent, DFMO, used to block the biosynthesis of polyamines. Many malignant cells can circumvent DFMO therapy by activating their transport system. A potential solution is to simultaneously block biosynthesis and transport of polyamines. However, little is known about the polyamine transport system in higher eukaryotes.

This thesis aims to add to the basic biological …

Pharmacology Of Organic Cation Transporters: Focus On Structure-Function Relationships In Oct3 (Slc22a3), Dan C. Li

Undergraduate Theses—Unrestricted

Organic Cation Transporters (OCTs) are polyspecific, facilitative transporters that play major roles in metabolite and drug clearance. OCTs are promising drug targets and elucidating their mechanisms of substrate recognition is crucial for rational drug design. OCT-mediated transport of polyvalent cations remains unexplored. OCT-expressing Xenopus laevis oocytes were used to assess transport of polyamines, ubiquitous polyvalent cations of broad physiological import, but for which transport mechanisms are unknown. Dose-response analysis of radiolabelled substrate uptake revealed that polyamines are relatively low affinity, but high turnover substrates for OCTs compared to model substrate methyl-4-phenylpyridinium (MPP⁺). Polyamine analogs of varying hydrophobic character …

Analyses Of Consumer Perceptions, Farmer Challenges, Polyamine, And Amino Acid Content Of Organic And Conventional Fresh Produce Grown In South Carolina, Emily Steinberg

All Dissertations

Execution of the USDA organic standards led to more support for local food as distinct from organic food. The current study was conducted to examine fresh produce perceptions and purchasing decisions of S.C. consumers. Four-hundred and eight S.C. consumers were surveyed at S.C. grocery stores. More than one-third of the S.C. consumers not knowing about the 'certified S.C. grown' program suggest that the S.C. Department of Agriculture (SCDA) may want to improve the promotion of their program. Eighty-five percent of consumers indicated that they would choose local over organic. This information would be useful to local S.C. produce farmers who …

Effect Of Water-Deficit Stress On Cotton During Reproductive Development, Dimitra Loka

Graduate Theses and Dissertations

Water deficit is a major abiotic factor limiting plant growth and crop productivity around the world. Cotton (Gossypium hirsutum L.) is considered to be relatively tolerant to drought and the effects of water stress on leaf physiology and metabolism have been extensively documented. However, information is lacking on the effect of water-deficit stress on the cotton flower. It was hypothesized that water-deficit stress would impair gas exchange functions which consequently would result in perturbation of carbohydrates of cotton reproductive units. To investigate this hypothesis growth room studies and field studies were conducted with the objectives being to document the physiological …

Investigating Cold Hardiness And Management Practices Of Warm-Season Putting Green Species In The Transition Zone, John M. Kauffman

Doctoral Dissertations

Warm-season turf species are becoming increasingly popular for putting green use in the transition zone. Ultradwarf bermudagrass (Cynodon dactylon (L.)  C. transvaalensis Burtt-Davy) is the prevalent warm-season putting green species, but seashore paspalum (Paspalum vaginatum Swartz) and ‘Diamond’ zoysiagrass [Zoysia matrella (L.) Merr.] may also be grown in the transition zone. Warm-season species are susceptible to winter injury and may require different management regimes than cool-season species. Therefore, the objectives of this research were to assess the impacts of various management practices on warm-season putting green species and characterize the physiological basis for differences in freeze tolerance of various …