Life Sciences Commons^™

Towards Understanding Osmolyte Effects On Folate(S) And Dihydrofolate Reductase Proteins, Purva Prashant Bhojane

Doctoral Dissertations

Osmolytes are small molecules that alter water activity and probe role of water in biological processes. Osmotic stress approach explored the role of water in ligand binding to dihydrofolate reductase (DHFR). DHFR catalyzes NADPH dependent reduction of dihydrofolate (DHF) to tetrahydrofolate (THF), which is essential for the synthesis of DNA, amino acids and other metabolic intermediates. R67 DHFR is a plasmid-encoded DHFR that confers resistance against trimethoprim, which is a potent inhibitor of E.coli chromosomal DHFR.

Osmolytes addition decreases the affinity of the substrate towards both the DHFRs. Weak preferential interactions between the osmolytes and DHF impede substrate binding to …

Spatial Crowding And Confinement Effects On Bursty Gene Expression, Charles Wei-Shing Chin

Doctoral Dissertations

Synthetic biology and genetic engineering are valuable tools in the development of new, sustainable energy generation technologies. The characterization of stochastic gene expression is vital to the efficient application of genetic engineering techniques. Transcriptional bursting, in which periods of high expression are punctuated by periods of no expression, is extensively observed in gene expression. While various molecular mechanisms have been hypothesized to be responsible for transcriptional bursting, spatial considerations have largely been neglected. This work uses computational modeling to examine in detail the influence of spatial factors such as macromolecular crowding and confinement on gene expression.

In the first part …

Tetrameric Photosystem I: From Initial Discovery And Characterization In Chroococcidiopsis Sp. Ts-821 To Exploration Of Its Distribution And Understanding Of Its Significance In Cyanobacteria, Meng Li

Doctoral Dissertations

Photosystem I (PSI) forms trimeric complexes in most characterized cyanobacteria. We had reported the tetrameric form of PSI in the unicellular cyanobacterium, Chroococcidiopsis sp. TS-821 (TS-821). Using Cryo-EM, a 3D model of the PSI tetramer structure at 11.5 [Angstrom] resolution was obtained and a 2D map within the membrane plane of at 6.1 [Angstrom]. In contrast to the three-fold symmetry in trimeric PSI crystal structure from T. elongatus, two different inter-monomer interactions involving PsaLs are found in the PSI tetramer. Phylogenetic analysis based on PsaL protein sequences shows that TS-821 is closely related to heterocyst-forming cyanobacteria. Additionally, this tetrameric …

Switchgrass Extractives Have Potential As A Value-Added Antimicrobial Against Plant Pathogens And Foodborne Pathogens, Alexander Ian Bruce

Masters Theses

Panicum virgatum (switchgrass), a perennial grass native to North America, is a leading biomass feedstock candidate for the manufacture of cellulosic ethanol. Switchgrass is considered a viable option for biofuel production due to its cheap production cost and ability to grow on marginal land. Biofuel derived from switchgrass has been shown to be very energy efficient, producing 540% more renewable energy versus nonrenewable energy expended. Switchgrass-derived biofuel is also estimated to have greenhouse gas emissions that are 94% lower than emissions from gasoline (Schmer et al 2008). Biofuels are created through biochemical processes that utilize various enzymes and microorganisms for …

Effect Of Varying Rumen Degradable And Undegradable Protein On Milk Production And Nitrogen Efficiency In Lactating Dairy Cows Under Summer Conditions, Jeffrey D. Kaufman

Masters Theses

The objective is to determine the effect of reducing nitrogen input through feeding low rumen degradable protein (RDP) and rumen undegradable protein (RUP) proportions on milk production, nitrogen efficiency and metabolism in heat-stressed cows. Forty-eight mid-lactating, Holstein cows were assigned to treatments using a randomized block design in a 2x2 factorial arrangement of treatments (n = 12/treatment). Treatments included two levels of RDP (10 and 8%) and two levels of RUP (8 and 6%). From d 1 to 21, a common diet (10% RDP-8% RUP) was fed to cows followed with their respective treatment diets fed from d 22 to …

Exploring Enrichment Cultures Of Denitrifying Microorganisms From El Yunque National Forest, Samiha Ahsan, Jenny Onley, Frank Loeffler

Chancellor’s Honors Program Projects

No abstract provided.

Characterization Of Fusarium Species And Alternaria Alternata, And Their Effects On Switchgrass Health And Chemical Components, Sara Beth Collins

Masters Theses

Panicum virgatum L., readily referred to as Switchgrass, is a perennial warm-season bunch grass, used as an alternative energy source for biofuel production. There is insufficient research on switchgrass pathogens, and is expected that an increase in disease pressure will result as more land is reserved for this perennial crop. The purpose of this research was to identify and characterize pathogenic Alternaria alternata and Fusarium species on Switchgrass (Panicum virgatum L.), and to evaluate their impact on stand establishment and plant health. Molecular identifications of fungal isolates obtained from infected commercial switchgrass seed yielded eight species (A. alternata, …

Rapid Molecular Detection And Population Genetics Of Pityophthorus Juglandis, A Vector Of Thousand Cankers Disease In Juglans Spp., Emel Oren

Masters Theses

Thousand Cankers Disease (TCD) is a disease complex involving the fungal pathogen Geosmithia morbida, an insect vector Pityophthorus juglandis, and the hosts, Juglans spp. and Pterocarya spp. Signs and symptoms of TCD include crown thinning due to branch dieback, yellowing and wilting of the leaves, appearance of epicormic shoots, numerous entrance/exit holes, gallery formation by P. juglandis, and the development of small, dark brown cankers underneath the bark. TCD originally described from western U.S., has now expanded to eastern U.S. and northwestern Italy. The disease complex is often difficult to diagnose due to the absence of symptoms …

Lignin Maximization: Analyzing The Impact Of Different Feedstocks And Feedstock Ratios Using Organosolv Fractionation, Marc Banholzer

Masters Theses

Over-exploitation of fossil fuels coupled with increasing pressure to reduce carbon emissions are prompting a transition from conventional petrochemical feedstocks to sustainable and renewable sourced carbon. The use of lignocellulosic biomass as a feedstock for integrated biorefining is of current high interest, as separation into its component parts affords process streams of cellulose, hemicellulose and lignin, each of which can serve as a starting point for the production of biobased chemicals and fuels. Given the large number of potential sources of lignocellulosic feedstocks, the biorefinery will need to adapt to the supplies available over a normal growing season. Of particular …

Molecular Dynamics Simulations Of Enzymes With Quantum Mechanical/Molecular Mechanical Potentials, Yufei Yue

Doctoral Dissertations

S-adenosyl methionine (SAM) dependent methylation process is universally found in all branches of life. It has important implications in mammalian pathogenesis and plant metabolism. The methyl transfer is normally catalyzed by SAM-dependent methyltransferases(MTases). Two MTases are studied in this dissertation: the 1,7-dimethylxanthine methyltransferase (DXMT) which involve in plant caffeine biosynthesis, and the protein arginine methyltransferase 5(PRMT5) that participates in eukaryotic posttranslational modification. The late phase of caffeine biosynthesis starts from the substrate xanthosine and ends with the product caffeine, with theobromine as an intermediate product. DXMT is a key enzyme in this process and catalyzes two methylation steps: 1)methylation of …

Electron Transport To Photosystem I By Soluble Carriers: Evolution Of The Interacting Pair, Khoa Dang Nguyen

Doctoral Dissertations

Oxygenic photosynthesis is driven via sequential action of Photosystem II (PSII) and (PSI) reaction centers via the Z-­‐scheme. Both of these pigment– membrane protein complexes are found in cyanobacteria, algae, and plants. PSI, unlike PSII, is remarkably stable and does not undergo limiting photo-­‐damage. This stability, as well as other fundamental structural differences, makes PSI the most attractive reaction centers for applied photosynthetic applications. These applied applications exploit the efficient light harvesting and high quantum yield of PSI where the isolated PSI particles are redeployed providing electrons directly as a photocurrent or, via a coupled catalyst to yield H2. Here, …

Modification Of Carbohydrate Active Enzymes In Switchgrass (Panicum Virgatum L.) To Improve Saccharification And Biomass Yields For Biofuels, Jonathan Duran Willis

Doctoral Dissertations

The natural recalcitrance of plant cell walls is a major commercial hurdle for plant biomass to be converted into a viable energy source as alternative to fossil fuels. To circumvent this hurdle manipulation of carbohydrate enzymes active in the cellulose and hemicellulose portions of the plant cell wall can be utilized to improve feedstocks. Production of cellulolytic enzymes by plants have been evaluated for reducing the cost associated with lignocellulosic biofuels. Plants have successfully served as bioreactors producing bacterial and fungal glycosyl hydrolases, which have altered plant growth to improve saccharification. A bioprospecting opportunity lies with the utilization of insect …

Decoding The Cellular Zipcode: Functional Analysis Of Transit Peptide Motifs And Mechanistic Implications In Plastid Targeting And Import, Kristen N. Holbrook

Doctoral Dissertations

Eukaryotic organisms are defined by their compartmentalization and various organelles. The membranes that define these organelles require complex nanomachines (known as translocons) to selectively mediate the import of proteins from the cytosol where they are synthesized into the organelle. The plastid, (specifically the chloroplast) which is characteristic of plant cells, possibly represents the most complex system of protein sorting, requiring many different translocons located in the three membranes found in this organelle. Despite having a small genome, the vast majority of plastid-localized proteins are nuclear-encoded and must be post-translationally imported from the cytosol. These proteins are encoded as a larger …

Characterization Of An Ethylene Receptor In Synechocystis Sp. Pcc 6803, Randy Francis Lacey

Doctoral Dissertations

In plants, ethylene functions as a hormone regulating many growth and developmental processes. Ethylene receptors in plants resemble bacterial two-component signaling systems. Because of this it, ethylene receptors are thought to have been acquired by gene transfer from the cyanobacterial endosymbiont that lead to the development of the chloroplast. However, prior to this work, functional ethylene receptors were thought to only be found in green plants. Here, we show that the cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis) contains a functional ethylene receptor, SynEtr1. SynEtr1 contains a predicted ethylene binding domain, a photosensory cyanobacteriochrome (CBCR) domain, and a histidine …

Nanocomposite Adhesive Of English Ivy (Hedera Helix): Bioproduction, Nanoparticle Isolation, And Molecular Analysis, Jason Neil Burris

Doctoral Dissertations

English ivy naturally produces organic nanoparticles from its adventitious root hairs, and possesses characteristics that may allow them to replace metal-based nanoparticles in common applications, such as sunscreen. At the onset of the research, it was hypothesized that a physical mechanism of attachment, similar to the gecko footpad, was used to generate the adhesive force for attachment; however, through the results obtained from recent work, it is clear that a biochemical mechanism is involved in the generation of the strength of adhesion. Therefore, the goal of this research was to provide a better understanding of the genetic basis of nanoparticle …

New Insights Into An Old Interaction: Developing A Model For Pai-1:Vn Interactions, Letitia Nichole Puster

Doctoral Dissertations

Active human Plasminogen Activator Inhibitor 1 (PAI-1) is most often found in complex with Vitronectin (VN), an ~62kDa glycoprotein. Research has shown PAI-1 and VN form higher order complexes in tissues, and our work indicates a 2:1 (PAI-1:VN) stoichiometry for these complexes. A logical model for PAI-1:VN interaction proposes that two PAI-1 molecules bind VN at separate sites. However, our small-angle neutron scattering (SANS) data suggest that there is a PAI-1: PAI-1:VN interaction, in which PAI-1 forms a dimer when in complex with VN. We tested this novel arrangement of PAI-1 within the complex by using a variety of biophysical …

Pinpointing The Molecular Basis For Metal Ion Effects On Plasminogen Activator Inhibitor-1 (Pai-1), Joel Cullen Bucci

Doctoral Dissertations

Plasminogen activator inhibitor type-1 (PAI-1) specifically inhibits the proteases tissue type plasminogen activator and urokinase plasminogen activator to control the activation of fibrinolysis. Vitronectin interacts with PAI-1 primarily through the somatomedin B (SMB) domain to stabilize and localize PAI-1 to sites of injury. Our laboratory observed that transition metals such as copper²⁺ have VN dependent, reciprocal effects on how long PAI-1 remains active. We aim to determine the molecular basis for effects of copper²⁺ on PAI-1 activity. We employed a computational algorithm (MUG) to predict metal binding clusters, and introduced mutations hypothesized to create metal binding deficiency. We …

Biophysical Studies Of Cell Division Protein Localization Mechanisms In Escherichia Coli, Matthew Wayne Bailey

Doctoral Dissertations

How nanometer-scale proteins position accurately within micron-scale bacteria has intrigued both biologists and physicists alike. A critical process requiring precise protein localization is cell division. In most bacteria, cell division starts with the self-assembly of the FtsZ proteins into filaments that form a ring-like structure encircling the cell at its middle, the Z-ring. The Z-ring is a scaffold for additional proteins that synthesize the lateral cell wall which separates the two daughter cells. If division planes are misplaced relative to bacterial chromosomes, also called nucleoids, daughter cells with incomplete genetic material can be produced. In Escherichia coli, research carried out …

In Silico Driven Metabolic Engineering Towards Enhancing Biofuel And Biochemical Production, Richard Adam Thompson

Doctoral Dissertations

The development of a secure and sustainable energy economy is likely to require the production of fuels and commodity chemicals in a renewable manner. There has been renewed interest in biological commodity chemical production recently, in particular focusing on non-edible feedstocks. The fields of metabolic engineering and synthetic biology have arisen in the past 20 years to address the challenge of chemical production from biological feedstocks. Metabolic modeling is a powerful tool for studying the metabolism of an organism and predicting the effects of metabolic engineering strategies. Various techniques have been developed for modeling cellular metabolism, with the underlying principle …

Complex Non-Equilibrium Structural Dynamics In Globular Proteins, Xiaohu Hu

Doctoral Dissertations

Internal structural motions in proteins are essential to their functions. In this present dissertation, we present the results from an extensive set of molecular dynamics simulations of three very different globular proteins and demonstrate that the structural fluctuations observed are highly complex, manifesting in non-ergodic and self-similar subdiffusive dynamics with non-exponential relaxation behavior. The characteristic time of the motion observed at a given timescale is dependent on the length of the observation time, indicating an aging effect. By comparing the simulation results to the existing single-molecule fluorescence spectroscopic data on other globular proteins, we found the characteristic relaxation time for …

Computational Perspective For Developing Bioluminescent Yeast Estrogen Screens For Environmental Toxicology, Jun Wang

Doctoral Dissertations

The impact of endocrine disruptive chemicals to human and wild life health has raised serious public health concerns through the past decades. To address this concern, much research was involved to develop tools for screening and assessing the hormonal potential of these compounds. Yeast bioluminescent bioreporter assay was one of the tools developed as the result of these past research endeavors. In this dissertation, a yeast bioluminescent bioreporter assay system was evaluated for the screening of endocrine disruptors from both experimental and computational perspectives. The yeast bioluminescent bioreporters were first standardized and applied in the comparative study of traditional activated …

Insulators: A “Safety Guard” For Genome Stability In Drosophila Melanogaster, Ran An

Doctoral Dissertations

Chromatin insulators are DNA-protein complexes that assist in the formation of chromatin loop structures by mediating long-range contacts between distant sites, which regulate proper organization of the chromatin fiber within the tridimensional space of the nucleus. It is considered that this function of insulators is required for the regulation of gene expression during development and in differentiated cells. This thesis focuses specifically in the Suppressor of Hairy wing [Su(Hw)] insulator of Drosophila and its associated proteins, and explores the possibility that chromatin insulators are not only significant for regulation of gene expression, but are also essential for DNA replication and …

Schizophrenia: Understanding The Madness, Robert W. Slaven

Chancellor’s Honors Program Projects

No abstract provided.

Effects Of Plant Root Exudates On Bacterial Chemotaxis, Quincy A. Banks

Chancellor’s Honors Program Projects

No abstract provided.

Modulating Neuronal Activity: Copper, Isoproterenol, And Beta-Blockers On The Brain, Veronica Go

Chancellor’s Honors Program Projects

No abstract provided.

Chloroplastic Protein Iojap Is Important For Cold-Acclimation In Arabidopsis Thaliana, Amanda Louise Carey

Chancellor’s Honors Program Projects

No abstract provided.

Targeting Cancer: The Ph-Responsive Binding And Insertion Of Roxy7, Kristen Rae Booth

Chancellor’s Honors Program Projects

No abstract provided.

Low Temperature Phenotype In Arabidopsis Thaliana Mutant Ijt1 Is Affected By Splicing, Connie Liu

Chancellor’s Honors Program Projects

No abstract provided.