Life Sciences Commons^™

Hsp70 And Hsp40 Functionally Interact With Soluble Mutant Huntingtin Oligomers In A Classic Atp-Dependent Reaction Cycle, Gregor P. Lotz, Justin Legleiter, Rebecca Aron, Emily M. Sontag, Shao-Yi Huang, Cheping Ng, Charles G. Glabe, Leslie M. Thompson, Paul J. Muchowski

Biological Sciences Faculty Research and Publications

Inclusion bodies of aggregated mutant huntingtin (htt) fragments are a neuropathological hallmark of Huntington disease (HD). The molecular chaperones Hsp70 and Hsp40 colocalize to inclusion bodies and are neuroprotective in HD animal models. How these chaperones suppress mutant htt toxicity is unclear but might involve direct effects on mutant htt misfolding and aggregation. Using size exclusion chromatography and atomic force microscopy, we found that mutant htt fragments assemble into soluble oligomeric species with a broad size distribution, some of which reacted with the conformation-specific antibody A11. Hsp70 associated with A11-reactive oligomers in an Hsp40- and ATP-dependent manner and inhibited their …

Tryptophan Anchored Peptides In Lipid Bilayer Membranes: Control Of Peptide Orientation And The Phase Behavior Of Cholesterol-Containing Ternary Lipid Mixtures, Johanna Maria Rankenberg

Graduate Theses and Dissertations

Model WALP peptides and "next generation" WALP-derived hydrophobic model peptides were employed to discover principles that govern protein-lipid interactions in biological membranes.

Ternary cholesterol-containing lipid mixtures were examined in the presence of WALP peptides of different lengths (acetyl-GWW(LA)_nLWWA-ethanolamide, with n between 3 and 8). Deuterium NMR spectra from labeled lipids reveal that WALP peptides may stabilize lipid ordered "raft" domains and therefore promote lipid phase separation, albeit to a minor extent. The results depend upon whether dioleolyl- or diphytanoyl-phosphatidylcholine is present as the fluid lipid component.

Several WALP peptides were modified to remove anchoring Trp residues from one …

Mechanism Of N-Methylation By The Trna M1g37 Methyltransferase Trm5., Thomas Christian, Georges Lahoud, Cuiping Liu, Katherine Hoffmann, John J Perona, Ya-Ming Hou

Department of Biochemistry and Molecular Biology Faculty Papers

Trm5 is a eukaryal and archaeal tRNA methyltransferase that catalyzes methyl transfer from S-adenosylmethionine (AdoMet) to the N(1) position of G37 directly 3' to the anticodon. While the biological role of m(1)G37 in enhancing translational fidelity is well established, the catalytic mechanism of Trm5 has remained obscure. To address the mechanism of Trm5 and more broadly the mechanism of N-methylation to nucleobases, we examined the pH-activity profile of an archaeal Trm5 enzyme, and performed structure-guided mutational analysis. The data reveal a marked dependence of enzyme-catalyzed methyl transfer on hydrogen ion equilibria: the single-turnover rate constant for methylation increases by one …

High Flow Air Sampling For Field Detection Using Gas Chromatography-Mass Spectrometry, Jacolin Ann Murray

Theses and Dissertations

The ability to rapidly detect and identify hazardous analytes in the field has become increasingly important. One of the most important analytical detection methods in the field is gas chromatography-mass spectrometry (GC-MS). In this work, a hand-portable GC-MS system is described that contains a miniature toroidal ion trap mass analyzer and a low thermal mass GC. The system is self-contained within the dimensions of 47 x 36 x 18 cm and weighs less than 13 kg. Because the instrument has a small footprint, it was used as the detector for an automated near-real-time permeation testing system. In permeation testing, materials …

Targeted Insertion Of Cysteine By Decoding Uga Codons With Mammalian Selenocysteine Machinery, Xue-Ming Xu, Anton A. Turanov, Bradley A. Carlson, Min-Hyuk Yoo, Robert A. Everley, Renu Nandakumar, Irina Sorokina, Steven P. Gygi, Vadim N. Gladyshev, Dolph L. Hatfield

Department of Biochemistry: Faculty Publications

Cysteine (Cys) is inserted into proteins in response to UGC and UGU codons. Herein, we show that supplementation of mammalian cells with thiophosphate led to targeted insertion of Cys at the UGA codon of thioredoxin reductase 1 (TR1). This Cys was synthesized by selenocysteine (Sec) synthase on tRNA^[Ser]Sec and its insertion was dependent on the Sec insertion sequence element in the 3′ UTR of TR1 mRNA. The substrate for this reaction, thiophosphate, was synthesized by selenophosphate synthetase 2 from ATP and sulfide and reacted with phosphoseryl-tRNA^[Ser]Sec to generate Cys-tRNA^[Ser]Sec. Cys was inserted in vivo at UGA …

Regulation Of Biofilm And Antibiotic-Resistance By The Modulator Of Sara (Msa) In Staphylococcus Aureus, Antony Schwartz

Dissertations

Staphylococcus aureus is an important human pathogen that is the causative agent of life-threatening diseases such as endocarditis and osteomyelitis. The ability of S. aureus to thrive as a successful pathogen can be partially attributed to its ability to form biofilm. Biofilm is an extracellular polysaccharide, protein, and DNA-based slime layer that protects the bacterial community. The global regulator sarA is essential for biofilm formation. Since the modulator of sarA (msa) gene regulates several virulence factors and is required for the full expression of sarA, the capacity of the msa mutant to form a biofilm was examined. The mutation of …

A Comprehensive Uncertainty Analysis And Method Of Geometric Calibration For A Circular Scanning Airborne Lidar, Michael Oliver Gonsalves

Dissertations

This dissertation describes an automated technique for ascertaining the values of the geometric calibration parameters of an airborne lidar. A least squares approach is employed that adjusts the point cloud to a single planar surface which could be either a narrow airport runway or a dynamic sea surface. Going beyond the customary three boresight angles, the proposed adjustment can determine up to eleven calibration parameters to a precision that renders a negligible contribution to the point cloud’s positional uncertainty.

Presently under development is the Coastal Zone Mapping and Imaging Lidar (CZMIL), which, unlike most contemporary systems that use oscillating mirrors …

Depth And Time Related Variations Of Microbial Communiites In An Emergent Freshwater Wetland, Amy Jenkins

Theses and Dissertations

Soils, and the microbial communities contained within them, are vital for most chemical, physical, and biological processes. This study investigated how microbial community structure responded to environmental changes, such as hydrology, across vertical space (depth) and time in an emergent fresh water wetland. Research was conducted in a non-tidal freshwater wetland along the James River (Charles City County, Virginia) by establishing plots in two areas that experienced different hydrologic regimes and plant communities. Soil cores (30 cm) were collected monthly from January 2008 to February 2009, and then every two to three months thereafter until October 2009, for a total …

Analysis Of Nitrogen Loading Reductions For Wastewater Treatment Facilities And Non-Point Sources In The Great Bay Estuary Watershed, Philip Trowbridge, Thomas S. Burack, Michael J. Walls, Harry T. Stewart

PREP Reports & Publications

In 2009, the New Hampshire Department of Environmental Services (DES) published a proposal for numeric nutrient criteria for the Great Bay Estuary. The report found that total nitrogen concentrations in most of the estuary needed to be less than 0.3 mg N/L to prevent loss of eelgrass habitat and less than 0.45 mg N/L to prevent occurrences of low dissolved oxygen. Based on these criteria and an analysis of a compilation of data from at least seven different sources, DES concluded that 11 of the 18 subestuaries in the Great Bay Estuary were impaired for nitrogen. Under the Clean Water …

Elucidating Functional Roles For Myogenin In Adult Skeletal Muscle Metabolism, Exercise Capacity, And Regeneration, Jesse Flynn

Dissertations and Theses (Open Access)

The four basic helix-loop-helix myogenic transcription factors, myogenin, Myf5, MRF4, and MyoD are critical for embryonic skeletal muscle development. Myogenin is necessary for the terminal differentiation of myoblasts into myofibers during embryogenesis, but little is known about the roles played by myogenin in adult skeletal muscle function and metabolism. Furthermore, while metabolism is a well-studied physiological process, how it is regulated at the transcriptional level remains poorly understood. In this study, my aim was to determine the function of myogenin in adult skeletal muscle metabolism, exercise capacity, and regeneration. To investigate this, I utilized a mouse strain harboring the Myogflox …

The Immune Inhibitor A1 Protease Of Bacillus Anthracis, Kathryn Pflughoeft

Dissertations and Theses (Open Access)

Bacillus anthracis, an organism ubiquitous in the soil and the causative agent of anthrax, utilizes multiple mechanisms to regulate secreted factors; one example is the activity of secreted proteases. One of the most abundant proteins in the culture supernates of B. anthracis is the Immune Inhibitor A1 (InhA1) protease. Here, I demonstrate that InhA1 modulates the abundance of approximately half of the proteins secreted into the culture supernates, including substrates that are known to contribute to the ability of the organism to cause virulence. For example, InhA1 cleaves the anthrax toxin proteins, PA, LF, and EF. InhA1 also targets …

Stress-Induced Targeting Of Molecular Chaperones In The Yeast Saccharomyces Cerevisiae, Hugo Tapia

Dissertations and Theses (Open Access)

The eukaryotic stress response is an essential mechanism that helps protect cells from a variety of environmental stresses. Cell death can result if cells are not able to properly adapt and protect themselves against adverse stress conditions. Failure to properly deal with stress has implications in human diseases including neurodegenerative disorders and distinct cancers, emphasizing the importance of understanding the eukaryotic stress response in detail. As part of this response, expression of a battery of heat shock proteins (HSP) is induced, which act as molecular chaperones to assist in the repair or triage of unfolded proteins. The 90-kDa HSP (Hsp90) …

Plasmacytoid Dendritic Cells Sense Skin Injury And Promote Wound Healing Through Type I Interferons, Josh D. Gregorio

Dissertations and Theses (Open Access)

Plasmacytoid dendritic cells (pDCs) are a rare population of circulating cells, which selectively express intracellular Toll-like receptors (TLR)-7 and TLR-9 and have the capacity to produce large amounts of type I IFNs (IFN-a/b) in response to viruses or host derived nucleic acid containing complexes. pDCs are normally absent in skin but accumulate in the skin of psoriasis patients where their chronic activation to produce IFN-a/b drives the disease formation. Whether pDCs and their activation to produce IFN-a/b play a functional role in healthy skin is unknown. Here we show that pDCs are rapidly and transiently recruited into healthy human and …

Tracking The Flow Of Information Through The Hippocampal Formation In The Rat, Joshua P. Neunuebel

Dissertations and Theses (Open Access)

The hippocampus receives input from upper levels of the association cortex and is implicated in many mnemonic processes, but the exact mechanisms by which it codes and stores information is an unresolved topic. This work examines the flow of information through the hippocampal formation while attempting to determine the computations that each of the hippocampal subfields performs in learning and memory. The formation, storage, and recall of hippocampal-dependent memories theoretically utilize an autoassociative attractor network that functions by implementing two competitive, yet complementary, processes. Pattern separation, hypothesized to occur in the dentate gyrus (DG), refers to the ability to decrease …

The Role Of Post-Translation Regulation Of Twist Expression In The Tumor Progression Of Squamous Cell Carcinoma Of Head And Neck, Ying-Wen Su

Dissertations and Theses (Open Access)

Squamous cell carcinoma of head and neck (SCCHN) is the tenth most common cancer in the world. Unfortunately, the survival of patients with SCCHN has not improved in the last 40 years. Therefore new targets for therapy are needed, and to this end we are studying signaling pathways activated by IL-6 which we have found stimulates cell migration and soft agar growth in SCCHN. Our data show that IL-6 increases TWIST expression in a transcription-independent mechanism in many SCCHN cell lines. Further investigation reveals TWIST can be phosphorylated upon IL-6 treatment. By computation prediction (http://scansite.mit.edu/motifscan_seq.phtml ), we found that TWIST …

Network To Pology In Human Protein Interaction Data Predicts Functional Association, Hua Li

Dissertations and Theses (Open Access)

High-throughput assays, such as yeast two-hybrid system, have generated a huge amount of protein-protein interaction (PPI) data in the past decade. This tremendously increases the need for developing reliable methods to systematically and automatically suggest protein functions and relationships between them. With the available PPI data, it is now possible to study the functions and relationships in the context of a large-scale network. To data, several network-based schemes have been provided to effectively annotate protein functions on a large scale. However, due to those inherent noises in high-throughput data generation, new methods and algorithms should be developed to increase the …

Characterization Of The Agrobacterium Tumefaciens Virb2 Pilin Of The Virb/D4 Type Iv Secretion System, Jennifer Kerr

Dissertations and Theses (Open Access)

The Agrobacterium tumefaciens VirB/D4 type IV secretion system (T4SS) delivers oncogenic T-DNA and effector proteins to susceptible plant cells. This leads to the formation of tumors termed Crown Galls. The VirB/D4 T4SS is comprised of 12 subunits (VirB1 to VirB11 and VirD4), which assemble to form two structures, a secretion channel spanning the cell envelope and a T-pilus extending from the cell surface. In A. tumefaciens, the VirB2 pilin subunit is required for assembly of the secretion channel and is the main subunit of the T-pilus. The focus of this thesis is to define key reactions associated with the T4SS …

Agrobacterium Virb10 Contributions To Type Iv Substrate Secretion, T-Pilus Biogenesis, And Outer Membrane Pore Formation, Isaac Garza

Dissertations and Theses (Open Access)

The VirB/D4 type IV secretion system (T4SS) of Agrobacterium tumefaciens functions to transfer substrates to infected plant cells through assembly of a translocation channel and a surface structure termed a T-pilus. This thesis is focused on identifying contributions of VirB10 to substrate transfer and T-pilus formation through a mutational analysis. VirB10 is a bitopic protein with several domains, including a: (i) cytoplasmic N-terminus, (ii) single transmembrane (TM) α-helix, (iii) proline-rich region (PRR), and (iv) large C-terminal modified β-barrel. I introduced cysteine insertion and substitution mutations throughout the length of VirB10 in order to: (i) test a predicted transmembrane topology, (ii) …

Analyzing The Role Of Αvβ8 Integrin In Glioma-Induced Angiogenesis, Jeremy H. Tchaicha

Dissertations and Theses (Open Access)

In this study we aimed to determine the functional roles for αvβ8 integrin in astrocytoma-induced angiogenesis. These studies originate from our analyses of αvβ8 integrin in developmental brain angiogenesis. αv and β8 knockout (KO) mice develop brain-specific vascular phenotypes that resemble vascular pathologies observed in the malignant astrocytoma, glioblastoma multiforme (GBM). Indeed, a murine xenograft model of astrocytoma suggested a role for the integrin in glioma-induced angiogenesis. Primary mouse astroglia were cultured from wild type (WT) and β8 KO neonates and were immortalized (HPV:E6/E7) and transformed (HRas:G12V). WT and β8 KO transformed astroglia were intracranially injected into athymic mice. WT …

14-3-3sigma Negatively Regulates The Stability And Subcellular Localization Of Cop1, Chun-Hui Su

Dissertations and Theses (Open Access)

Mammalian constitutive photomorphogenic 1 (COP1), a p53 E3 ubiquitin ligase, is a key negative regulator for p53. DNA damage leads to the translocation of COP1 to the cytoplasm, but the underlying mechanism remains unknown. We discovered that 14-3-3σ controlled COP1 subcellular localization and protein stability. Investigation of the underlying mechanism suggested that, upon DNA damage, 14-3-3σ bound to phosphorylated COP1 at S387, resulting in COP1 translocation to the cytoplasm and cytoplasmic COP1 ubiquitination and proteasomal degradation. 14-3-3σ targeted COP1 for degradation to prevent COP1-mediated p53 degradation, p53 ubiquitination, and p53 transcription repression. COP1 expression promoted cell proliferation, cell transformation, and …

Dynamic Remodeling Of The Stressed Heart: Role Of Protein Degradation Pathways, Deborah Vela

Dissertations and Theses (Open Access)

The heart is a remarkable organ. In order to maintain its function, it remodels in response to a variety of environmental stresses, including pressure overload, volume overload, mechanical or pharmacological unloading and hormonal or metabolic disturbances. All these responses are linked to the inherent capacity of the heart to rebuild itself. Particularly, cardiac pressure overload activates signaling pathways of both protein synthesis and degradation. While much is known about regulators of protein synthesis, little is known about regulators of protein degradation in hypertrophy. The ubiquitin-proteasome system (UPS) selectively degrades unused and abnormal intracellular proteins. I speculated that the UPS may …

Mcam/Muc18 Regulates Melanoma Progression By Modulating The Expression Of Id-1, Maya Zigler

Dissertations and Theses (Open Access)

The acquisition of the metastatic melanoma phenotype is associated with increased expression of the melanoma cell adhesion molecule MCAM/MUC18 (CD146). However, the mechanism by which MUC18 contributes to melanoma metastasis remains unclear. Herein, we stably silenced MUC18 expression utilizing lentivirus-incorporated small hairpin RNA, in two metastatic melanoma cell lines, A375SM and C8161, and conducted cDNA microarray analysis. We identified and validated that the transcriptional regulator, Inhibitor of DNA Binding-1 (Id-1), previously shown to function as an oncogene in several malignancies, was downregulated by 5.6-fold following MUC18 silencing. Additionally, we found that MUC18 regulated Id-1 expression at the transcriptional level via …

4-H Leaders: Factors That Affect Their Persistence In The 4-H Youth Development Program, Regina Schinker

Dissertations

For over one hundred years, the 4-H youth development program has educated millions of children across the country in a variety of programs; from livestock husbandry to leadership skills. Like all programs that have endured, 4-H has had to adjust its focus as society has changed. One area in the 4-H program that has not changed is the heavy reliance on adult volunteer leaders to deliver the educational programs to 4-H youth. Because of the reliance on volunteer 4-H leaders, 4-H professionals and extension office staff are concerned about issues of retention.

The case study examines two related issues: First, …

Identification And Characterization Of Micrornas In Porcine Gametes And Pre-Implantation Embryos, Erin Curry

All Dissertations

MicroRNAs (miRNAs) are short ribonucleic acids that ultimately affect the production of proteins. Although miRNAs are involved in nearly every biological process examined to date, little is known of the identity or function of miRNA in porcine reproductive tissues or their potential involvement in reproductive processes in pigs or other species. The objective of this dissertation research was to determine the presence of miRNAs in porcine gametes and both in vivo- and in vitro- produced pre-implantation embryos and to identify differences in miRNA expression between normal and aberrant samples. Using a heterologous RT-PCR approach, we demonstrated the presence of a …

Resistance And Dna Repair Mechanisms Under Nitrosative Stress In Mammalian And Microbial Systems, Hyun-Wook Lee

All Dissertations

Living organisms are exposed a nitrosative stress mediated by reactive nitrogen species (RNS) that can cause DNA damage and mutation. DNA base deamination is a typical damage occurred under nitrosative stress, which results in conversion of cytosine (C) to uracil (U), adenine (A) to hypoxanthine (I), and guanine (G) to xanthine (X) or oxanine (O). Base excision repair (BER) is an important pathway to remove deaminated DNA lesions in mammalian and microbial systems. My dissertation work concerns with genes and enzymes involved in resistance to nitrosative stress and DNA glycosylases in the BER pathway. In chapter one, I will briefly …

Population Variability Of Rotylenchulus Reniformis In Cotton Agroecosystems, Megan Leach

All Dissertations

Rotylenchulus reniformis, reniform nematode, is a highly variable species and an economically important pest in many cotton fields across the southeast. Rotation to resistant or poor host crops is a prescribed method for management of reniform nematode. An increase in the incidence and prevalence of the nematode in the United States has been reported over the last two decades. However it is not clear whether the observed increase is related to the emergence of novel populations that are more aggressive or have a higher fitness or shifts in host availability or susceptibility. The objectives of this research were to determine …

Development And Characterization Of Novel Sub-Coatings With Polymorphic Brookite Titania Nanoparticles: Enhanced Uv/Vis Photocatalytic Antibacterial And Anticancer Properties, Donna Weinbrenner

All Dissertations

ABSTRACT
Nosocomial and community infections and biofilm formation from bacteria has increased significantly through adaptation combined with overuse of broad spectrum antibiotics. Because the world population continues to escalate, hospitals and long-term managed care will also escalate, thereby increasing transmission of infections, lowering patients' quality of health. Another disease on the rise throughout the world is skin cancer. A treatment modality that would cause less deleterious effects on the patient would be ideal. These two seemingly different issues could be solved with one product. First an inexpensive, safe, and non-selective antimicrobial surface coating would enhance the hospital arena and a …

Sequencing And Functional Analysis Of A Multi-Component Dioxygenase From Pah-Degrading Sphingomonas Paucimobilis Epa505, Renuka Miller

All Dissertations

Polycyclic aromatic hydrocarbons (PAHs) are hydrophobic organic compounds consisting of two or more fused benzene rings. PAHs derive from many different sources including petroleum refining, wood treatment, and coal coking industries. Because of their structural stability and water insolubility, PAHs are extremely resistant to degradation. These compounds are also believed to have mutagenic, carcinogenic, and teratogenic effects. Therefore, there are currently 16 PAH compounds on the EPA's list of priority pollutants.
Many species of bacteria have the ability to breakdown these persistent pollutants. However, bioremediation strategies using these organisms have many unresolved issues. While laboratory experiments can easily demonstrate the …

A New Partial Skeleton Of A Cryptocleidoid Plesiosaur From The Upper Jurassic Sundance Formation Of Wyoming, Benjamin C. Wilhelm, F. Robin O’Keefe

Biological Sciences Faculty Research

Cryptocleidoid plesiosaurs from the Upper Jurassic are well known from the Oxford Clay (Callovian) of the United Kingdom. The plesiosaurs of the nearly coeval Sundance Formation (Oxfordian) of North America are poorly known, but are thought to include two cryptocleidoid taxa:Pantosaurus striatus and Tatenectes laramiensis. Here we present two specimens recently recovered from the Bighorn Basin of Wyoming. The first specimen comprises three articulated adult cervical vertebrae and fragments of a fourth. This specimen preserves a posteriorly directed cervical neural spine, a character diagnostic of Pantosaurus striatus. It also resembles Pantosaurus in the morphology of its cervical …

Current Distribution Of Rare Fishes In Eastern Wyoming Prairie Streams, Christina E. Barrineau, Elizabeth A. Bear, Anna C. Senecal

The Prairie Naturalist

Distributions of native fishes have declined throughout the Great Plains region. Over 50% of native fishes within the Missouri River drainage in Wyoming have experienced declines in distributions. Thus, the primary goal of our study was to assess current distribution of rare native fishes in eastern Wyoming prairie streams. Of the 10 rare fishes sampled, goldeye (Hiodon alosoides), western silvery minnow (Hybognathus argyritis), plains minnow (H. hankinsoni), and Iowa darter (Etheostoma exile) have experienced declines in distribution over the last decade. Plains topminnow (Fundulus sciadicus) appears to be expanding to areas outside …