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Non-Natural Nucleotides As Probes For The Mechanism And Fidelity Of Dna Polymerases, Irene Lee, Anthony J. Berdis 2010 Case Western Reserve University

Non-Natural Nucleotides As Probes For The Mechanism And Fidelity Of Dna Polymerases, Irene Lee, Anthony J. Berdis

Chemistry Faculty Publications

DNA is a remarkable macromolecule that functions primarily as the carrier of the genetic information of organisms ranging from viruses to bacteria to eukaryotes. The ability of DNA polymerases to efficiently and accurately replicate genetic material represents one of the most fundamental yet complex biological processes found in nature. The central dogma of DNA polymerization is that the efficiency and fidelity of this biological process is dependent upon proper hydrogen-bonding interactions between an incoming nucleotide and its templating partner. However, the foundation of this dogma has been recently challenged by the demonstration that DNA polymerases can effectively and, in some ...


Physiological And Molecular Function Of Hap3b In Flowering Time Regulation And Cold Stress Response, Mingxiang Liang 2010 Utah State University

Physiological And Molecular Function Of Hap3b In Flowering Time Regulation And Cold Stress Response, Mingxiang Liang

All Graduate Theses and Dissertations

Heme-activated proteins (HAPs) are transcription factors that have multiple roles in plant growth and development, such as embryogenesis, flowering time control, and drought tolerance. In the present study I found that HAP3b was also involved in controlling response to cold stress. Transcript profiling and gene expression analyses indicated that HAP3b repressed the CBF3 regulon under normal growth conditions. As a result, plants with HAP3b-overexpressed showed decreased survival rates while plants homozygous for the null allele hap3b showed an improved freezing tolerance compared to wild-type plants. To understand the mechanism of HAP3b in Arabidopsis, i.e. whether it also acts through ...


Role Of Protein Kinase C Isotypes In 1,25-Dihydroxyvitamin D3 Mediated Signal Transduction Through The 1,25d3 Membrane Associated, Rapid Response Steroid Binding Receptors In Chick Intestinal Cells, Sakara Tunsophon 2010 Utah State University

Role Of Protein Kinase C Isotypes In 1,25-Dihydroxyvitamin D3 Mediated Signal Transduction Through The 1,25d3 Membrane Associated, Rapid Response Steroid Binding Receptors In Chick Intestinal Cells, Sakara Tunsophon

All Graduate Theses and Dissertations

It is now accepted that 1,25(OH)2D3 mediates its rapid actions on the control of phosphate and calcium homeostasis through its membrane receptor termed the 1,25D3-MARRS (membrane associated rapid response steroid binding) protein. I determined the various PKC isotypes involved in the rapid regulation of phosphate uptake and calcium extrusion in chick intestinal cells. 1,25(OH)2D3-mediated phosphate uptake was stimulated within 1 min after addition of the hormone. Western blot analyses on isolated intestinal cells treated with steroid hormone resulted in dose-dependent increases in PKC alpha and PKC beta in postnuclear centrifugation fractions, but not ...


Determining The Role Of Water And Proton Uptake/Release Upon Binding Of Substrate Dhf And Cofactor Nadph To The Enzyme Dihydrofolate Reductase's Active Site, Sharghi Rahmanian 2010 University of Tennessee - Knoxville

Determining The Role Of Water And Proton Uptake/Release Upon Binding Of Substrate Dhf And Cofactor Nadph To The Enzyme Dihydrofolate Reductase's Active Site, Sharghi Rahmanian

Chancellor’s Honors Program Projects

No abstract provided.


Circadian Expression Patterns Of Proteins In The Suprachiasmatic Nucleus, Jenna E. McKinnie 2010 University of Tennessee - Knoxville

Circadian Expression Patterns Of Proteins In The Suprachiasmatic Nucleus, Jenna E. Mckinnie

Chancellor’s Honors Program Projects

No abstract provided.


Regulation Of The Mitotic Cyclin Clb2 In The Budding Yeast Saccharomyces Cerevisiae, Cody R. Richardson 2010 University of Tennessee - Knoxville

Regulation Of The Mitotic Cyclin Clb2 In The Budding Yeast Saccharomyces Cerevisiae, Cody R. Richardson

Chancellor’s Honors Program Projects

No abstract provided.


Functional Analysis Of An Alternative Replication Protein A Complex Containing Rpa4, Aaron Charles Mason 2010 University of Iowa

Functional Analysis Of An Alternative Replication Protein A Complex Containing Rpa4, Aaron Charles Mason

Theses and Dissertations

Replication Protein A (RPA), the eukaryotic single-stranded DNA-binding complex, is essential for multiple processes in cellular DNA metabolism including, but not limited to, DNA replication, DNA repair and recombination. The `canonical' RPA is composed of three subunits (RPA1, RPA2, and RPA3). In addition to the three canonical subunits, there is a human homolog to the RPA2 subunit, termed RPA4, which can substitute for RPA2 in complex formation. The resulting RPA complex has been termed `alternative' RPA (aRPA). The normal function of aRPA is not known; however, previous studies have shown that it does not support S-phase progression in vivo. The ...


Bioluminescent Mouse Models Of Prostate Cancer Progression And Therapy, Robert Ulf Svensson 2010 University of Iowa

Bioluminescent Mouse Models Of Prostate Cancer Progression And Therapy, Robert Ulf Svensson

Theses and Dissertations

Prostate cancer is the most common visceral neoplasm and second leading cause of cancer death in US men. It is a complex heterogeneous and multi-factorial disease whose mechanistic basis is poorly understood. Furthermore, treatment options for advanced metastatic prostate cancer are limited and do not impart significant survival benefits, highlighting the need for novel and more effective therapeutic strategies. To this end we examined the use of RNAi therapy based on the systemic delivery of optimized small interfering RNAs (siRNAs) targeting the androgen receptor (AR). siRNA treatment induced massive cell death in vitro but was unable to induce tumor regression ...


Enzymatic Regulation Of Phosphatidylcholine Synthesis Via Protein Ubiquitination, Phillip Louis Butler 2010 University of Iowa

Enzymatic Regulation Of Phosphatidylcholine Synthesis Via Protein Ubiquitination, Phillip Louis Butler

Theses and Dissertations

Pulmonary surfactant is a critical surface-active substance consisting of dipalmitoylphosphatidylcholine (DPPtdCho) and key apoproteins that are produced and secreted into the airspace from alveolar type II epithelial cells. Deficiency of the surfactant leads to severe lung atelectasis, ventilatory impairment, and gas-exchange abnormalities. The generation of DPPtdCho in cells occurs via two integral routes: the de novo and remodeling pathways. The interplay between these pathways has not been investigated. Overexpression of the remodeling enzyme, acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT1), in epithelia decreases de novo PtdCho synthesis without significantly altering cellular phospholipid mass; this occurs through increased degradation of cholinephosphotransferase (CPT1), the terminal ...


Micrornas' Role In Brain Development And Disease, Sarah Kathryn Fineberg 2010 University of Iowa

Micrornas' Role In Brain Development And Disease, Sarah Kathryn Fineberg

Theses and Dissertations

MicroRNA (miRNA) function is required for normal animal development, in particular in stem cell and precursor populations. I hypothesize that miRNAs are similarly required for stem cell maintenance and appropriate fate commitment in the brain. To test the requirement for global microRNA production, I depleted the microRNA biosynthetic enzyme DICER in the developing mouse brain. I found that DICER loss in embryonic neural progenitor cells leads to embryonic lethality with microcephaly. By histological analysis, I found defects in both neural progenitor cell maintenance and cell differentiation. I also identified new candidate microRNAs for this phenotype by profiling miRNAs in DICER-depleted ...


Host Signaling Response To Adhesion Of Bifidobacterium Infantis, Reed N. Gann 2010 Utah State University

Host Signaling Response To Adhesion Of Bifidobacterium Infantis, Reed N. Gann

All Graduate Theses and Dissertations

Investigations of the molecular binding partners of the probiotic bacterium Bifidobacterium longum subspecies infantis (B. infantis) and the pathogen Salmonella enterica subspecies enterica serovar Typhimurium LT2 (Salmonella ser. Typhimurium) found that these two very different bacteria bind gangliosides. However, these organisms lead to completely different host health outcomes when present in the gut. B. infantis is the founding microbial population in the intestinal tract of breast-fed infants. S. typhimurium is the most important food-borne pathogen that results in humans. This study used an in vitro gut epithelial cell model to examine the host cellular response to adhesion of B. infantis ...


Transesterification And Recovery Of Intracellular Lipids Using A Single Step Reactive Extraction, Daniel R. Nelson 2010 Utah State University

Transesterification And Recovery Of Intracellular Lipids Using A Single Step Reactive Extraction, Daniel R. Nelson

All Graduate Theses and Dissertations

A single-step, extractive reaction for extraction of lipids such as biodiesel components, omega-3 fatty acids, or other triglycerides from microbial cells was examined. Conventional methods for lipid extraction use toxic solvents, and require multiple steps and long processing times. When the goal is to produce fatty acid methyl esters or FAMEs, the extracted lipids are subjected to a separate transesterification reaction with simple alcohols in the presence of an acid or base catalyst. A simplified, single-step reactive extraction method can be applied that combines the sequential extraction followed by transesterification using acidified alcohols - a process known as in situ transesterification ...


High-Throughput Screening Studies Of Inhibition Of Human Carbonic Anhydrase Ii And Bacterial Flagella Antimicrobial Activity, Albert A. Barrese III 2010 Western Michigan University

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.


Antimicrobial Activity Of D-Lenolate®, Andy Phui 2010 University of Nevada Las Vegas

Antimicrobial Activity Of D-Lenolate®, Andy Phui

UNLV Theses, Dissertations, Professional Papers, and Capstones

Olive trees are one of the most important fruit trees in the Mediterranean. Although not validated by research, olive leaves are traditionally believed to fight off fever and infections. It has been shown that olive leaf extracts possess antimicrobial activity. Olive leaf extracts contain polyphenols. One of the major phenolic compounds is oleuropein. Oleuropein and other polyphenols have been shown to exhibit antimicrobial activity. East Park Research (EPR) developed an extraction process that they claim does not alter the chemical composition of the olive leaves. The extract is known by the commercial name d-lenolate®. Studies have provided evidence that d-lenolate ...


Development Of Novel Methods And Their Utilization In The Analysis Of The Effect Of The N-Terminus Of Human Protein Arginine Methyltransferase 1 Variant 1 On Enzymatic Activity, Protein-Protein Interactions, And Substrate Specificity, Brenda Bienka Suh-Lailam 2010 Utah State University

Development Of Novel Methods And Their Utilization In The Analysis Of The Effect Of The N-Terminus Of Human Protein Arginine Methyltransferase 1 Variant 1 On Enzymatic Activity, Protein-Protein Interactions, And Substrate Specificity, Brenda Bienka Suh-Lailam

All Graduate Theses and Dissertations

Protein arginine methyltransferases (PRMTs) are enzymes that catalyze the methylation of protein arginine residues, resulting in the formation of monomethylarginine, and/or asymmetric or symmetric dimethylarginines. Although understanding of the PRMTs has grown rapidly over the last few years, several challenges still remain in the PRMT field. Here, we describe the development of two techniques that will be very useful in investigating PRMT regulation, small molecule inhibition, oligomerization, protein-protein interaction, and substrate specificity, which will ultimately lead to the advancement of the PRMT field. Studies have shown that having an N-terminal tag can influence enzyme activity and substrate specificity. The ...


Carboxylases Involved In Microbial Acetone And Acetophenone Metabolism, Ameya Mashruwala 2010 Utah State University

Carboxylases Involved In Microbial Acetone And Acetophenone Metabolism, Ameya Mashruwala

All Graduate Plan B and other Reports

A number of bacteria are capable of growth with acetone and acetophenone as their sole sources of carbon and/or energy. The pathways and enzymes involved in the transformation of these molecules into useable carbon and energy are unique. Among these are two novel enzymes, acetone carboxylase and acetophenone carboxylase, which represent a fundamentally novel classes of carboxylases.

The initial step in acetone metabolism, in X. autotrophicus st Py2, R. capsulatus st B10 and R. rhodochrous, is the thermodynamically unfavorable reaction to yield acetoacetate. This step is catalyzed by the enzyme acetone carboxylase and is coupled with the unprecedented, concomitant ...


Bacillus Cereus And Bacillus Anthracis Germination Kinetics: A Michaelis-Menten Approach, Helen Luu 2010 University of Nevada, Las Vegas

Bacillus Cereus And Bacillus Anthracis Germination Kinetics: A Michaelis-Menten Approach, Helen Luu

UNLV Theses, Dissertations, Professional Papers, and Capstones

Bacillus species are rod-shaped, gram-positive bacteria that are capable of producing endospores. In this dormant stage, the endospores can persist in hostile physical and chemical environments. Once conditions become favorable, the spores germinate into actively dividing cells, vegetative cells. Germination is a crucial step for the pathogenicity of the Bacilli in affecting a host organism.

Our study applies mathematical approaches to spore germination to determine whether the binding of one germinant will affect the binding of another germinant. We pursued this approach with two different species, B. cereus and B. anthracis, both pathogenic organisms. B. cereus is a widely known ...


A Synthetic Biological Engineering Approach To Secretion- Based Recovery Of Polyhydroxyalkanoates And Other Cellular Products, Elisabeth Linton 2010 Utah State University

A Synthetic Biological Engineering Approach To Secretion- Based Recovery Of Polyhydroxyalkanoates And Other Cellular Products, Elisabeth Linton

All Graduate Theses and Dissertations

The costs associated with cellular product recovery commonly account for as much as 80% of the total production expense. As a specific example, significant recovery costs limit commercial use of polyhydroxyalkanoates (PHA), which comprise a class of microbially-accumulated polyesters. PHAs are biodegradable compounds that are of interest as a sustainable alternative to petrochemically-derived plastics. Secretion-based recovery of PHAs was studied to decrease PHA production costs. Type I and II secretory pathways are commonly used for the translocation of recombinant proteins out of the cytoplasm of E. coli. Proteins were targeted for translocation using four signal peptides (HlyA, TorA, GeneIII, and ...


Mtor In Cell Signaling And Size Enlargement As A Target For The Chemico-Physical Therapy Of Cancer, Gabriela Krawiec 2010 Syracuse University

Mtor In Cell Signaling And Size Enlargement As A Target For The Chemico-Physical Therapy Of Cancer, Gabriela Krawiec

Syracuse University Honors Program Capstone Projects

mTOR is a kinase protein meaning it phosphorylates target proteins affecting their cell signaling properties1. The drug Rapamycin, analogs of Rapamycin, and cell signaling proteins that interact with mTOR control the activities mediated by mTOR1. mTOR is located in the cytoplasm at a convergent point of many signaling pathways that regulate a multiplicity of cellular processes including metabolism that precede cell enlargement (cell “growth”), cell proliferation (cell division), and angiogenesis1,2. Cells with mTOR inappropriately activated can proceed with cell enlargement and cell proliferation in the absence of normal cell signaling2. Rapamycin and Rapamycin analogs can ...


The Effect Of Exogenous Estrogens On Primordial Follicle Assembly In Vivo, Jenna Rose Karavan 2010 Syracuse University

The Effect Of Exogenous Estrogens On Primordial Follicle Assembly In Vivo, Jenna Rose Karavan

Syracuse University Honors Program Capstone Projects

During embryogenesis in the mouse, primordial germ cells develop, move to the genital ridge of the embryo, and form germline cysts as the ovary develops. The cells in these cysts are linked by intercellular bridges. The cysts then undergo a breakdown process which ultimately results in primordial follicles, each of which consists of a single oocyte surrounded by somatic cells called granulosa cells. During this cyst breakdown process, approximately one-third of the original oocytes become enclosed in primordial follicles, the amount of which is representative of the number of eggs a female will have available to her during her reproductive ...


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