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The Role Of The Mcm2 Subunit In Regulating The Activities Of The Mcm2-7 Helicase, Brent E. Stead

Electronic Thesis and Dissertation Repository

The transmission of genetic information from parental to daughter cells requires the faithful duplication of an organism’s genome. Uncontrolled DNA replication can result in proliferative diseases, such as cancer. DNA replication requires a single-stranded DNA template to be produced from duplex DNA. In eukaryotes, DNA unwinding for replication is performed by the heterohexameric replicative helicase complex comprised of the minichromosome maintenance proteins 2 through 7.

Each of the Mcm2-7 subunits likely has a unique role in DNA binding and unwinding by the Mcm2-7 complex. The present study examines the role of the Saccharomyces cerevisiae Mcm2 subunit in regulating the activities …

Identification Of Regions Responsible For The Open Conformation Of S100a10 Using Chimaeric S100a11/S100a10 Proteins, Liliana Santamaria-Kisiel

Electronic Thesis and Dissertation Repository

S100A11 is a dimeric, EF-hand calcium-binding protein. Calcium binding to S100A11 results in a large conformational change that uncovers a broad hydrophobic surface used to interact with phospholipid-binding proteins (annexins A1 and A2), and facilitate membrane vesiculation events. In contrast to other S100 proteins, S100A10 is unable to bind calcium due to deletion and substitution of calcium-ligating residues. Despite this, calcium-free S100A10 assumes an “open” conformation that is very similar to S100A11 in its calcium-bound state (Ca2+-S100A11). To understand how S100A10 is able to adopt an open conformation in the absence of calcium, seven chimeric proteins were constructed where regions …

Perception Meets Action: Fmri And Behavioural Investigations Of Human Tool Use, Kenneth F. Valyear

Electronic Thesis and Dissertation Repository

Tool use is essential and culturally universal to human life, common to hunter-gatherer and modern advanced societies alike. Although the neuroscience of simpler visuomotor behaviors like reaching and grasping have been studied extensively, relatively little is known about the brain mechanisms underlying learned tool use.

With learned tool use, stored knowledge of object function and use supervene requirements for action programming based on physical object properties. Contemporary models of tool use based primarily on evidence from the study of brain damaged individuals implicate a set of specialized brain areas underlying the planning and control of learned actions with objects, distinct …

Effects Of Acute Synchronous Whole-Body Vibration Exercise, Tom J. Hazell

Electronic Thesis and Dissertation Repository

Recently, whole-body vibration (WBV) exercise has been gaining interest in the health/fitness community for its reported beneficial outcomes. However, most of these have not been evaluated scientifically leading to some reservation in promoting this new exercise modality. The purpose of this dissertation was to assess the viability of WBV exercise to enhance several selected indices of health.

Study 1 demonstrated that the addition of WBV to an isometric semi-squat in young healthy men (n = 8, 25±2.6 y, 177±7.0 cm, 84±12.1 kg) resulted in increases in femoral artery blood flow and leg skin temperature vs the same exercise without vibration …

Structural Insights Into Dna Replication And Lesion Bypass By Y Family Dna Polymerases, Kevin N. Kirouac

Electronic Thesis and Dissertation Repository

Y family DNA polymerases are specialized enzymes for replication through sites of DNA damage in the genome. Although the DNA damage bypass activity of these enzymes is important for genome maintenance and integrity, it is also responsible for DNA mutagenesis due to the error-prone nature of the Y family. Understanding how these enzymes select incoming nucleotides during DNA replication will give insight into their role in cancer formation, aging, and evolution. This work attempts to mechanistically explain, primarily through X-ray crystallography and enzymatic activity assays, how Y family polymerases select incoming nucleotides in various DNA replication contexts. Initially, we sought …

Regulation Of Akt And Wnt Signalling By The Dopamine D2 Receptor And Metabotropic Glutamate Receptor 2/3, Laurie P. Sutton

Electronic Thesis and Dissertation Repository

Akt and the Wnt pathway, two cascades that regulate GSK-3, have been implicated in schizophrenia and antipsychotic drug action. Although it is known that antipsychotic drugs alleviate psychosis by blocking the dopamine D2 receptor (D2DR) and that metabotropic glutamate receptor 2/3 (mGluR2/3) agonists may improve some of the symptoms of schizophrenia, it is unclear if both classes of drugs exert their effects through Akt, GSK-3 and/or the Wnt pathway or if changes in these pathways are mediated through the D2DR and mGluR2/3 respectively. In addition to antipsychotics, mood stabilizers and antidepressants also target GSK-3, suggesting that there must be something …

Cardiac Adaptation To Chronic Blockade Of Voltage-Gated, L-Type Calcium Channels In The Sarcolemma, Ji Zhou

Electronic Thesis and Dissertation Repository

L-type Ca²⁺ channels (dihydropyridine receptors, DHPRs) in the sarcolemma are essential to cardiac excitation-contraction (E-C) coupling. Thus, Ca²⁺ influx through DHPRs upon cardiomyocyte excitation triggers Ca²⁺ release from the sarcoplasmic reticulum (SR) through ryanodine receptors (RyRs) to initiate myofilament activation and muscle contraction. Muscle relaxation occurs upon sequestration of Ca²⁺ back into the SR lumen by sarco/endoplasmic reticulum calcium-ATPase (SERCA) in the SR. As a treatment option for hypertension, long-term use of DHPR blockers is associated with increased risk of heart failure; underlying mechanisms are unknown. This research used male Wistar rats treated with verapamil (subcutaneously, …

Mental Blocks: The Behavioural Effects And Neural Encoding Of Obstacles When Reaching And Grasping, Craig S. Chapman

Electronic Thesis and Dissertation Repository

The ability to adeptly interact with a cluttered and dynamic world requires that the brain simultaneously encode multiple objects. Theoretical frameworks of selective visuomotor attention provide evidence for parallel encoding (Baldauf & Deubel, 2010; Cisek & Kalaska, 2010; Duncan, 2006) where concurrent object processing results in neural competition. Since the end goal of object representation is usually action, these frameworks argue that the competitive activity is best characterized as the development of visuomotor biases. While some behavioural and neural evidence has been accumulated in favour of this explanation, one of the most striking, yet deceptively common, demonstrations of this capacity …

Pharmacokinetics And Therapeutic Uses Of Mesna, Murray J. Cutler

Electronic Thesis and Dissertation Repository

In the early 1980s, significant advancement in the safety of ifosfamide therapy was achieved by co-administrating mesna (sodium 2-mercaptoethane sulfonate) to prevent dose-limiting hemorrhagic cystitis. Mesna exerts its protective effect within the urine, where its free sulfhydryl group is able to conjugate cytotoxic metabolites. Within the circulation, however, mesna exists primarily as its inactive disulfide, dimesna. Dimesna is currently undergoing clinical development as a prodrug (BNP7787) to treat cisplatin-induced nephrotoxicity. Remarkably, chemoprotection is achieved without attenuation of efficacy of co-administered anti-cancer agents. This is widely attributed to the kidney-specific disposition and stability of dimesna.

We sought to evaluate the role …

Stopover Biology Of Migratory Landbirds In A Heavily Urbanized Landscape, The New York Metropolitan Area, Chad L. Seewagen

Electronic Thesis and Dissertation Repository

Migration routes of many Nearctic-Neotropical landbirds pass through the most urbanized regions of North America. Migrants use urban habitat fragments as stopover sites and commonly occur in cities at exceptional density. Yet, knowledge of migrant stopover biology and refueling opportunities in such places is severely limited. This dissertation examined several aspects of migrant stopover biology in the New York metropolitan area to gain a more holistic understanding of how migratory landbirds utilize urban stopover sites, and ultimately to assess the quality of urban habitats as stopover sites. I first generated morphometric predictive models using salvaged bird specimens to allow me …

The Lotus Japonicus Cytokinin Receptor Gene Family And Its Role In Nitrogen-Fixing Symbiosis, Mark A. Held

Electronic Thesis and Dissertation Repository

Nitrogen is the most abundant element in our atmosphere, yet has become increasingly limited in agricultural lands. Legume plants offer a possible solution to this problem due to their innate ability to symbiotically interact with nitrogen-fixing bacteria called rhizobia. In particular, a histidine kinase cytokinin receptor from the model legume Lotus japonicus (LHK1) has been clearly placed at the core of these interactions. Loss-of-function mutants in LHK1 fail to initiate timely cortical cell divisions in response to abundant bacterial infection, and gain-of-function mutations in the same locus cause L. japonicus plants to form spontaneous nodules in the absence of rhizobia, …

Mitotic Regulation Of Protein Kinase Ck2, Nicole A. St. Denis

Electronic Thesis and Dissertation Repository

Protein kinase CK2 is a serine/threonine kinase with a multitude of substrates and roles in many cellular processes, including mitosis. CK2 is constitutively active, yet we hypothesize that CK2 is indeed regulated in mitosis through subtle means, enabling CK2 to perform its functions unique to cell division. Our aims were to examine the roles of mitotic phosphorylation, subcellular localization, and interplay with mitotic kinases in the regulation of CK2 activity.

We first examined the role of four highly conserved mitotic phosphorylation sites located in the unique C-terminus of CK2α. Phosphospecific antibodies generated against the sites show that CK2α phosphorylation is …

The Roles Of Nitric Oxide Synthases (Nos) In Endochondral Bone Formation, Qian Yan

Electronic Thesis and Dissertation Repository

Longitudinal growth of endochondral bones is controlled by the cartilage growth plate. Chondrocyte proliferation and hypertrophy, vascular invasion, formation of ossification centers and cartilage replacement by bone tissue are all important processes required for normal growth. These biological processes have to be tightly regulated or disturbances will lead to skeletal diseases. A large number of genes, growth factors and hormones have been implicated in the regulation of growth plate biology, however, less is known about the intracellular signaling pathways involved. Nitric oxide (NO) has been identified as a regulator of cellular proliferation, differentiation, migration, survival and metabolism in multiple cell …

Regulation Of Dna Damage Processing By Covalent Modification Of Thymine Dna Glycosylase, Ryan D. Mohan

Electronic Thesis and Dissertation Repository

Thymine DNA glycosylase (TDG) is an essential DNA repair enzyme mediating excision of uracil and thymine mispaired with guanine within CpG contexts. Unrepaired, these lesions result in G:C to A:T transitions which are major contributors to genome instability. Interestingly, TDG interacts functionally with transcriptional regulators and participates in directed cytosine demethylation at promoters. TDG is subject to multiple post-translational modifications (PTM) and we undertook an analysis of how these regulate TDG function. Initially, we examined TDG regulation by small ubiquitin-like modifier (SUMO) and identified a novel SUMO binding motif (SBM1, residues 144-148). We hypothesized that SBM1, along with SBM2 (319-322), …