Biochemistry, Biophysics, and Structural Biology Commons^™

The Binding Properties And Functional Consequences Of Ryr2-Cam Interaction, Yi Yang

Dissertations

The aim of my dissertation is to understand the regulation of RyR2. The whole dissertation is composed of two parts. The first part focused on RyR2-CaM interaction. The second focused on synthetic RyR2 domain peptide (DPc10), which worked as a powerful molecular tool for RyR2 functional and structural studies.

CaM has been long identified as an important cardiac RyR regulator. Broad studies suggest CaM is a critical RyR2 stabilizer and CaM-RyR2 interaction is a critical molecular substrate for arrhythmias and HF pathogenesis, but the in situ binding properties for CaM-RyR2 are still unknown. Here we, Using FRET detection and permeabilized …

Role Of Notch Signaling In T Cell Polarization, Shilpa Keerthivasan

Dissertations

The differentiation of CD4+ T cells to different effector lineages in response to pathogenic stimuli is the core of the adaptive immune system. One of the effector subsets recently discovered is Thelper 17 (Th17) and it plays a predominant role in autoimmune diseases and inflammatory disorders.

In my thesis, I aimed to study the role of Notch cell surface receptors in Th17 differentiation. Using in vitro Th17 differentiation assays of human naïve CD4+ T cells, I have shown that Notch signaling, particularly Notch1, plays a crucial role in Th17 polarization. By using pharmacological inhibitors and specific knockdown of Notch1, I …

Repression Of Protein Kinase C Delta In Human Squamous Cell Carcinomas By Ras, Fyn And Nf-Kappa B Signaling, Vipin Yadav

Dissertations

The delta isoform of Protein Kinase C (PKC-delta) is widely expressed in many normal tissues, including epidermal keratinocytes, and has a critical role in UV-induced apoptosis. However, PKC-delta is frequently lost in chemically or UV-induced mouse skin tumors, as well as in human cutaneous squamous cell carcinomas (SCC). Furthermore, re-expression of PKC-delta in human SCC lines is sufficient to induce apoptosis and suppress tumorigenicity, making PKC-delta a potential tumor suppressor gene for SCCs. The objective of this dissertation is to investigate the mechanism of PKC-delta loss in human SCCs.

To determine the mechanism of PKC-delta loss in human SCCs, we …

Glucosamine-Induced Insulin Resistance In Primary Rat Hepatocytes And The Role Of Selenium As An Insulin Mimetic, Sandhya N. Adiyodi Veetil

Dissertations

Type 2 diabetes is mediated by insulin resistance, the inability of insulin to elicit a normal biological response in insulin responsive tissues. Several cellular models have been utilized to determine the mechanism of induction of insulin resistance but questions remain unanswered. One model, implicates the products of the Hexosamine Biosynthetic Pathway (HBP) in the induction of insulin resistance under hyperglycemia. The major end product of HBP, UDP-GlcNAc, is the substrate for O-GlcNAc transferase, an enzyme that catalyzes the O-GlcNAcylation of numerous proteins. This modification may play a role in induction of insulin resistance and thus needs to be evaluated in …

Molecular, Cellular And Systemic Effects Of Atrazine On The Xenopus Laevis Tadpole, Renee Maryanne Zaya

Dissertations

Atrazine is one of the two most commonly used herbicides in the US and it is the most studies are among the first to link physiological effects of atrazine to changes in tissue and gene expression in Xenopus laevis tadpoles. These studies provide the groundwork for future studies into the mechanisms behind responses to chemical stressors generating data resulting in a better understanding of how these chemicals affect us all. commonly found herbicide in ground water. As a result, a great deal of attention has been placed on its use and safety. It has also been implicated to play a …

The Specific Role Of The Mll Cxxc Domain In Mll Fusion Protein Function, Laurie Ellen Risner

Dissertations

The MLL gene was first identified because it is involved in chromosome translocations which produce novel fusion proteins that cause leukemia. The CXXC domain of MLL is a cysteine rich DNA binding domain with specificity for binding unmethylated CpG-containing DNA. The CXXC domain is retained in oncogenic MLL fusions, and is absolutely required for the fusions to cause leukemia. This project explored the role of the CXXC domain by introducing structure-informed point mutations within the MLL CXXC domain that disrupt DNA binding, and by performing domain swap experiments in which different CXXC domains from other proteins, including DNMT1, CGBP and …

Role Of Non-Coding Rna Nc4 In Mll And Cyp33 Mediated Regulation Of Hoxc8, Jessica Arvindbhai Solanki

Dissertations

MLL or Mixed Lineage Leukemia gene is clinically known for its involvement in genetic translocation with more than 70 different partners identified each giving rise to a highly leukemogenic fusion protein. With a poor prognosis of MLL related leukemia, an investigation into its role during hematopoiesis has been a very active field of research. In order to design strategies to combat the MLL leukemia, it becomes essential to delineate the molecular mechanisms behind the function of MLL wild type protein. Wild type MLL is a transcriptional maintenance protein from the Trithorax Group (TrxG) that resides in complex with other chromatin …

High-Throughput Screening Studies Of Inhibition Of Human Carbonic Anhydrase Ii And Bacterial Flagella Antimicrobial Activity, Albert A. Barrese Iii

Dissertations

Complete abstract attached as supplemental file.

The Role Of Igf-1 And Notch Signaling In Thoracic Malignancies., Sandra Eliasz

Dissertations

Thoracic malignancies are one of the deadliest of all cancers, being the leading cause of cancer death in the Western world. Thoracic malignancies arise from different tissues; however the most common are of epithelial (commonly referred to as non-small cell lung cancer, or NSCLC), neuroendocrine (small cell lung cancer, or SCLC) and mesothelial origin (malignant mesothelioma, or MM). The DNA oncogenic virus Simian Virus 40 (SV40) has been shown to cooperate with environmental oncogenic fibers in the onset of MM. Insulin like growth factor-1 (IGF-1) signaling plays a central role in all thoracic malignancies and in the process of SV40-mediated …

New Cyclophanes As Supramolecular Scaffolds: The Synthesis Of Tribenzo-1,4,7-Triazacyclononatriene., Andria M. Panagopoulos

Dissertations

Supramolecular chemistry involves the formation of complex molecular entities that have the capacity to participate in specific molecular recognition of guest molecules. A commonly employed scaffold in supramolecular chemistry is the trimeric crown-shaped molecule cyclotriveratrylene (CTV). CTV has been studied extensively for its capability of binding a number of smaller organic and organometallic guests within its bowl-shaped cleft and has been used as a building block enabling the construction of more complex cryptophanes. The goal of this research is the synthesis and characterization of a novel cyclophane, tribenzo-1,4,7-triazacyclononene and derivatives thereof. These new cyclophanes should have greater versatility than the …

Polyomavirus Enhancer Activator 3 (Pea3), A Member Of The Ets Family Of Transcription Factors, Is A Transcriptional Activator Of Notch-1 And Notch-4 In Breast Cancer: An Opportunity For Novel Combinational Therapy, Anthony George Clementz

Dissertations

Women diagnosed with triple-negative breast cancer have the worst overall prognosis and frequently present with metastatic tumors. To date, there are no targeted therapies available to combat this aggressive form of breast cancer due to the lack of expression of well-known targets such as ER-alpha, PR, or HER2/neu. Therefore, there is an immediate need to identify novel targets that are responsible for the proliferation, survival, and invasive phenotype. Notch-1 and Notch-4, both potent breast oncogenes, are overexpressed in triple-negative breast cancers-associated with the poorest overall survival. PEA3 (polyomavirus enhancer activator 3), a member of the Ets family of transcription factors, …

Developmental Characterization Of Cara Mitad: A Drosophila Nuclear Receptor Co-Regulator, Chhavi Chauhan

Dissertations

In insects and vertebrates, hormone titers drive cellular proliferation and differentiation events that guide proper development. Nuclear receptors (NR) respond to these hormone signals by activating cascades of gene expression, along with coregulator protein complexes. In Drosophila melanogaster, fluctuating titers of the steroid hormone ecdysone are responsible for coordinating the timing of organismal development. Despite major advances in our understanding of Drosophila NR activities, we lack essential knowledge of the coregulators that are required for their proper function.

We have recently identified the Drosophila cara mitad (cmi) (`dear half') gene. The deduced CMI protein is closely related to the N-terminal …

Lysine Specific Demethylase-1 And The Brahma Chromatin Remodeling Complex Regulate Conserved Signaling Pathways During Drosophila Wing Development, Brenda Jean Curtis

Dissertations

The conserved SWI/SNF chromatin remodeling complex uses the energy from ATP hydrolysis to alter local chromatin environments by disrupting DNA-histone contacts. The Drosophila SWI/SNF counterpart, the Brahma complex, has been shown to have an essential role in regulating the proper expression of many developmentally important genes, including those required for eye and wing tissue morphogenesis. A temperature sensitive mutation in one of the core subunits, SNR1 (SNF5/Ini1/SMARC B1), results in reproducible wing patterning phenotypes that can be enhanced and suppressed by extragenic mutations. SNR1 functions as a regulatory subunit to modulate chromatin remodeling activities of the Brahma complex on target …

Biochemical Characterization Of Two Yeast Paralogous Proteins Mth1 And Std1, Satish Pasula

Dissertations

Glucose is the most abundant monosaccharide and preferred carbon and energy source for most cells. Many organisms have evolved sophisticated means to sense glucose and respond to it appropriately. The budding yeast, Saccharomyces cerevisiae senses glucose through two transmembrane proteins, Snf3 and Rgt2. In the presence of extracellular glucose Snf3 and Rgt2 generate an intracellular signal that leads to the degradation of Mthl and Stdl, thereby inducing the expression of hexose transporter genes (EXT) by inhibiting the function of Rgtl, a transcriptional repressor of HXT genes. Mthl and Stdl are degraded via the Yckl/2 Kinase-SCFGrrl-26S proteasome pathway triggered by the …

The Effects Of Aging And Activity On Glial Cell Line-Derived Neurotrophic Factor Expression In Skeletal Muscle, Nathan Peplinski

Dissertations

Glial cell line-derived neurotrophic factor (GDNF) is an extracellular signaling protein that is produced by skeletal muscle and is important for the motor neurons that control muscle movement. GDNF has been shown to keep neurons alive under conditions that they would otherwise not persist. In skeletal muscle, GDNF has been shown to be one of the most potent neurotrophic factors that influence motor neuron survival. While the role of GDNF has been well studied during early development, not much is known about what happens to GDNF expression in the adult and with advanced aging. Previous results from our lab have …

Structure-Function Studies Of Self- Assembling Flagellin Proteins Fromsalmonella Typhimurium And Aquifex Pyrophilus, Venkata Raghu Ram Malapaka

Dissertations

The eubacterial flagellum is a complex structure with an elongated extracellular filament that is primarily composed of a single protein termed flagellin. The highly conserved N-and C- termini of flagellin are important in its export and assembly, whereas the middle "hypervariable" region is highly variable in size across different species and is largely dispensable. InSalmonella typhimurium phase 1 flagellin (FliC), this hypervariable region encodes two solvent-exposed D2 and D3 domains. The functional role of this knob-like structural feature in motility remains unclear. The structural and physiological role of the hypervariable region in flagellar assembly, stability and cellular motility was investigated. …

Characterization Of Molecular Events Following Heavy Metal Cadmium Exposure: A Cellular Regulatory Symphony In Response To Early Cadmium Insult, Chin-Ju Jean Hsiao

Dissertations

Cadmium, a non-essential toxic heavy metal, has been classified as a human carcinogen and one of the most hazardous substances in our environment. Since it can circulate within the food chain, has a long biological half-life and can bio-accumulate, it has the potential to induce detrimental health effects linked to toxicity, cancer and chronic diseases.

In recent years progress has been made to dissect and understand the molecular mechanisms underlying the effects of cadmium exposure. However, few studies have focused on investigating potential protective and/or defense mechanisms naturally applied by cells upon initial exposure, prior to any damage or toxic …

The Glucose And Insulin Responsiveness Of The Rat Glucose-6-Phosphate Dehydrogenase Promoter, Daryl Arkwright-Keeler

Dissertations

The regulation of expression of some enzymes involved in glucose homeostasis and fat metabolism requires both insulin and glucose to elicit an effect, while the regulation of others requires only one. Glucose-6-phosphate dehydrogenase (G6PDH), an important enzyme involved in glucose homeostasis, has been shown to be transcriptionally regulated by carbohydrate feeding in rat liver, a diet regime that increases the levels of both glucose and insulin. Our lab had previously shown that insulin induces expression of G6PDH, thus we wanted to determine if glucose also induced G6PDH expression and if so, are the glucose and insulin responses mediated through identical …

Bufadienolides In The Chemical Defenses Of The Toads, Bufo Americanus And Bufo Fowleri, Catherine E. Merovich

Dissertations

I investigated the steroidal chemical defenses (bufadienolides) of Bufo americanus and Bufo fowleri. By the nature of their complex, biphasic life cycles, toads, like other amphibians are important components of aquatic and terrestrial habitats and are prey to numerous invertebrates and vertebrates. Bufadienolides are presumed to be important anti-predatory compounds although much of their chemical ecology is poorly understood. I investigated (1) ontogenetic variation in bufadienolides, (2) bufadienolides from adult parotoid secretion, (3) effectiveness of bufadienolides against a terrestrial predator, and (4) effectiveness of bufadienolides against an aquatic predator. I hypothesized that B. americanuswould have a more extensive …

Health Impacts Of Polychlorinated Biphenyls (Pcbs) In Aquatic Organisms, Marla Ann Fisher

Dissertations

Polychlorinated biphenyls (PCBs) are a major contaminant of global extent in water resources and aquatic biota. Due to its high lipid solubility, PCBs fail to be degraded and, therefore, continue to bioaccumulate throughout the environment and food chain. To determine the impact of PCBs on the immune system ofaged and juvenile Japanese medaka (Oryzias latipes), fish were injected with the coplanar PCB congener 126 and examined after 3 and 14 days. PCB 126 produced oxidative stress in both age groups of fish 14 days post-injection; however, juvenile medaka appeared more susceptible than aged fish. Humoral immunity, as determined by antibody …

K+- Induced Dilation Of Cremasteric Arterioles Involves Na+/K+Atpase And Inward Rectifier K+ Channels, Wendy Burns

Dissertations

The purpose of this study was to evaluate the possible mechanisms involved in cremasteric arteriolar dilation as a result of increased concentrations of ([K⁺]_o) extracellular potassium. Many other studies suggested that Na⁺/K⁺ ATPase or K_IR channels are responsible for potassium-induced vasodilation in other systems (2, 12, 22, 40, 46, 52, 16, 17, 25, 33, 34, 85, 99, 151). However, these studies show only partial inhibition of dilation as a result of independent blockageof each mechanism. The hypothesis of this study was that both mechanisms contribute to potassium-induced dilation seen in cremasteric arterioles. …

Molecular Staging Of Colorectal Cancer: K-Ras Mutation Analysis Of Lymph Nodes Upstages Dukes’ B2 Patients, Jennifer S. Thebo

Dissertations

The successful treatment of colorectal cancer depends upon the ability of staging systems to identify patients who are at risk for recurrence. The currently used systems fail in a significant number of patients. Multiple attempts have been made to improve upon these systems in order to provide better treatment for at-risk patients.

A variety of oncogenes and tumor suppressor genes have been implicated in colorectal cancer. Mutations in one of these, K-ras, occur in 40-60% of colorectal carcinomas. However, the relationship between specific mutations and clinical outcome is unclear.

The purpose of this study was to determine whether or not …

The Electrophysiological Effects Of Acute Thyroid Hormone Exposure On Atrial Myocytes, Jon Paul Fiening

Dissertations

No abstract provided.

Genetic Analysis Of The Nuo Locus That Encodes The Proton Translocating Nadh Dehydrogenase In Escherichia Coli, Holly Jennifer Falk-Krzesinski

Dissertations

No abstract provided.

Singlet-Oxygen-Mediated Inactivation Of Acetylcholinesterase: A Comparison Of Purified Enzyme In Solution And Enzyme Bound To K562 Leukemia Cells, Gail D. Deadwyler

Dissertations

No abstract provided.

Analysis Of The Mechanisms Controlling Phospholipid Biosynthetic Gene Expression In The Yeast Saccharomyces Cerevisiae, Brian Patrick Ashburner

Dissertations

No abstract provided.

Developmental, Tissue, And Sub-Tissue Specific Expression Of The Mammalian Apurinic/Apyrimidinic Endonuclease, Teresa Mari Wilson

Dissertations

No abstract provided.

Regulation Of Phospholipid Biosynthesis In The Yeast Saccharomyces Cerevisiae By The Ume6 Gene, John Charles Jackson

Dissertations

No abstract provided.

Cloning Of Multiple Novel Human Trinucleotide Repeat Containing Cdna's: A Novel Application Of Rapid Amplification Of Cdna Ends (Race), James P. Carney

Dissertations

No abstract provided.

Analysis Of Amino Acids Relevant To Activity And Substrate Specificity Of The Human Dna Repair Protein 0-6-Methylguanine Dna Methyltransferase (Mgmt), Susan Elizabeth Morgan

Dissertations

No abstract provided.