Digital Commons Network^™

Novel Role Of Endoplasmic Reticulum-Associated Degradation In The Regulation Of Ceruloplasmin, Stephen William Hippleheuser

Wayne State University Theses

Ceruloplasmin (Cp) is a secreted ferroxidase produced by the hepatocytes that assists in the transport of iron throughout the human body. In human aceruloplasminemia patients, due to Cp deficiency excess intracellular iron buildup leads to ailments like liver cirrhosis, neurodegeneration, and blindness. We recently found that the biogenesis of Cp in the endoplasmic reticulum (ER) is regulated by a principal ER quality-control process, ER-associated degradation (ERAD). ERAD clears misfolded ER proteins for cytosolic proteasomal degradation, with the Sel-1 suppressor of lin-12-like (Sel1L)-HMG-CoA reductase degradation 1 (Hrd1) protein complex representing the most conserved branch in mammals. Interestingly, we found that Sel1L-Hrd1 …

Determination Of The Direct Protein-Protein Interactions In The Drosophila Sin3a Complex, Ian Moore

Wayne State University Theses

The proteins that comprise the Drosophila SIN3 220 and SIN3 187 complexes are currently known. Limited information with regard to interacting complex member proteins has been described. Much of these data are results of high-throughput investigations, and there have been no studies done to reconstitute all direct interactions within the complex. The unique C-terminal region of the SIN3 220 isoform was used to test for interactions with other complex proteins using the bacterial expression system. Additionally, the region of Caf1-55 necessary for interaction with SIN3 in vivo was identified using a truncation mutant. The results of this work identified novel …

Synaptotagmin C2b Ca2+-Binding Loops Impose Distinct Exocytosis Phenotypes, Michael W. Schmidtke

Wayne State University Theses

Regulated exocytosis from chromaffin cells in the adrenal medulla plays a critical role in maintaining organismal homeostasis. In the absence of stress, these cells release physiologically relevant substances into the blood stream only in limited quantities, whereas stressful conditions result in a rapid deluge of signaling molecules used, for example, to increase heart rate and pain tolerance. Although the cellular mechanisms governing the switch from low-level to stress-induced secretion are not well understood, recent evidence has implicated the exocytotic Ca2+-sensing protein Synaptotagmin (Syt) in this role.

Two isoforms of Syt are expressed in chromaffin cells (Syt-1 and Syt-7), and each …

Development Of A Dreissena Bioassay To Assess The Toxicity Of Contaminants Across Two Life-History Stages, Caroline Joyce Addis

Wayne State University Theses

Dreissenid mussels (Dreissena polymorpha and D. bugensis) have rapidly become widespread and ubiquitous in North America since their introduction into the Great Lakes in the 1980s. The resulting environmental and economic impacts of their invasion have been extensive, negatively affecting biodiversity and costing millions of dollars in control efforts and damage to power generation and water treatment facilities. Although dreissenids are often associated with negative impacts, they may present a practical tool for toxicology studies. The typically sessile behavior of the benthic adults coupled with the planktonic nature of the veligers allow for a more complete evaluation of water quality …

Functional Characterization Of The Novel Prdm Gene Apox In Tribolium Castaneum, Xin Wang

Wayne State University Theses

Previous work revealed that embryonic and post-embryonic knockdown (KD) of Tribolium Prdm gene Apoptix (Apox) leads to phenotypes, such as melanotic spots in external tissue of larval and pupal body. One part of my research is screening for Apox-specific phenotypes by RNA interference with a second Apox dsRNA. My results revealed that Apox is essential for the embryonic and post-embryonic survival and plays an important role in the regulation of embryonic and post-embryonic development in Tribolium, such as bristle formation. In addition, previous work showed that phenotypes of Apox KD Tribolium were significantly rescued by combinatorial knockdown of Apox and …

Novel Biochemical And Phenotypical Findings On The Nodal Signaling Components Lefty And Cripto, Amapola Balancio

Wayne State University Theses

ABSTRACT

NOVEL BIOCHEMICAL AND PHENOTYPICAL FINDINGS ON THE NODAL SIGNALING COMPONENTS LEFTY AND CRIPTO

by

Amapola Balancio

December 2014

Advisor: Dr. William Branford, Jr.

Major: Biological Sciences (Molecular and Developmental Biology)

Degree: Master of Science

Nodal, a Transforming Growth Factor-β (TGF-B), is a key regulator of proper development along with its atypical TGF-B inhibitor Lefty and its EGF-CFC co-receptor Cripto. Studies have shown that Nodal is unable to signal in the absence of a functional Cripto, indicating that it is a Nodal obligate co-receptor. In fact, one of the ways that Lefty prevents Nodal is by binding to Cripto and …

The Contribution Of The Melanin Pathway To Overall Body Pigmentation Changes During Ontogenesis Of Periplaneta Americana, Thomas Roger Lemonds

Wayne State University Theses

Pigmentation is one of the most diverse and distinguishable features of insect morphology. The most prominent colors observed in insects are black or brown, whose production is attributed to the melanin pathway. At present, though, the contribution of this pathway to overall body pigmentation throughout ontogenesis is still lacking. To address this question we examined the roles of four key melanin genes (TH, DDC, ebony, and aaNAT), in embryonic and post-embryonic development of Periplaneta americana. Our results show show that while the melanin pathway plays no role during the earliest nymphal stages, it is required during the later stages of …

Acoustic Cues Of Physical Formidability In Cage Fighters, Stefan Mattias-Maria Goetz

Wayne State University Theses

Across the animal kingdom, the sex that experiences the most reproductive variance tends to evolve sexually dimorphic traits—both behavioral and morphological—which aid in reproduction. Human evolution has been marked by greater male intrasexual selection and as a result, men display a variety of secondary sexual characteristics, putatively serving to enhance biological fitness. Among these, fundamental frequency, closely related to perception of pitch, among men is half that of women. Likewise, monotonicity, that is, variance in pitch across an utterance, is higher in men (i.e., women show greater variance in pitch) while formant dispersion, which gives the voice its timbre, is …

The Effects Of Phragmites Australis Litter On Seed Emergence In The Erie-Huron Corridor, Michigan, Travis White

Wayne State University Theses

The invasive reed, Phragmites australis, is widespread within the Great Lakes region, and is often blamed for habitat degradation. Once established, it creates dense litter mats that may persist following remediation efforts of living stock removal. We investigated the effects of P. australis and Typha angustifolia, narrow-leaf cattail, litter on seedling emergence from the native seed bank by harvesting soils from five Great Lakes coastal marshes densely populated by either Phragmites or Typha and exposing them to Phramities or Typha litter in treatments of varying litter depths. Seedling emergences were quantified for six weeks. Soils from Phragmites dominated sites had …

The Phenotypic Analysis Of The Knockdown Of The Sin3a Complex Components And Their Role In Recruitment And Cell Proliferation, Kelly Ann Laity

Wayne State University Theses

The SIN3A-RPD3 complex components have previously been identified in Drosophila melanogaster. The role of these components in SIN3A function and recruitment was not known. Polytene chromosome analysis following RNAi knockdown was performed to determine if any of the complex members affect the ability of SIN3A to bind to chromatin. The complex components effect on cell proliferation was also examined through clonal analysis of imaginal wing discs. The results of this work implicate a role of several members of the SIN3A complex for proper recruitment and localization to chromatin. All of the SIN3A complex members had some varying effect on cell …

Synthesis And Characterization Of Brain Penetrant Prodrug Of Neuroprotective D264: Potential Disease Modifying Treatment Agent For Parkinson's Disease, Fahd Shamoon Dholkawala

Wayne State University Theses

Parkinson's disease (PD) is a complex neurodegenerative disorder with progressive loss of dopamanergic neurons in the substantia nigra region of the brain and accumulation of intracytoplasmic inclusions called `Lewy bodies'. PD is characterized by tremors, rigidity, slowness of movement, bradykinesia and postural imbalances. Although the etiology of PD is not well understood, it is well established that oxidative stress, mitochondrial dysfunction, alpha-synuclein aggregation play a central role in the pathogenesis of PD. Current treatment methods are based on symptomatic relief without addressing the underlying pathophysiological factors responsible for the disease. It is important to develop therapies which can address these …

Approaches To Improving Detection Of Invasive Fish Species In Western Lake Erie Through Analysis Of Monitoring Efficiencies And Metrics Of Community Distribution, Joshua Alan Southern

Wayne State University Theses

Efficient monitoring programs are essential for the early detection of invasive species. The Ohio Department of Natural Resources (ODNR) monitoring program encompassing 21 years of fish survey data from western Lake Erie was evaluated using Chao biodiversity analysis to determine the efficiency and precision of collection strategies of trawl and gillnet sampling, at detecting rare or non-native species. Overall, ODNR sampling annually accounted for ~80% of extant fish species, leaving gaps in coverage where rare and invasive species may be overlooked and proliferate.Obtaining 90% efficiency would require an estimated doubling of previous sampling effort. Computer simulations calculating different proportions of …

Role Of Phosphoylation Of The Mycobacterium Tuberculosis Proteasome In Oxidative Stress Response And Regulation Of Homoserine Kinase (Thrb) In Corynebacterium Glutamicum, Heather Baun

Wayne State University Theses

Part 1:

One-third of the world's population is infected with the latent form of Mycobacterium tuberculosis. M. tuberculosis must be able to control this latent even in response to nitrosative and oxidative stress conferred by the host. It is not well known how M. tuberculosis controls this state.

One possibility for this control is through its eleven eukaryotic-like serine/threonine protein kinases. Our lab has focused research on two eukaryotic-like ser/thr protein kinases, PknA and PknB. It has been found that PknA expression may affect the stability or the assembly of the proteasome complex, which is essential for M. tuberculosis to …

Defining The Mechanism Of Enhanced Cellular Invasion Induced By Mechanical Stimulation, Snehal Sunil Ozarkar

Wayne State University Theses

Metastasis is a multistep process driven by various biochemical and mechanical factors, which eventually leads to formation of secondary tumors. The tumor mass is surrounded by basement membrane (BM) and stroma made of various extracellular matrix (ECM) proteins. During metastasis tumor cells disseminate from the primary tumor, breach the BM, invade the stroma, travel through blood and lymph and colonize tissues distant from the primary tumor. Formation of secondary tumors by metastasis is a leading cause of death in cancer patients. Even though plenty of research has been focused on biochemical factors affecting metastasis, information on role of mechanical factors …

Landscape Characteristics Of Upland Sandpiper Habitat In Michigan, Jacob Lawrence Korte

Wayne State University Theses

Grassland bird populations have declined across North America due to habitat loss but at a disproportionately higher rate in the midwestern United States, where extensive coverage of grasslands and other open land ecosystems have been converted to other land cover types. The upland sandpiper (Bartramia longicauda Bechstein, UPSA) is a migratory, area-sensitive, terrestrial shorebird that breeds in grasslands and other open land ecosystem types across their North American range. Although breeding habitats of the Great Plains contain the greatest concentrations of this species, anthropogenic openings such as hayfields and pastures serve as surrogate habitat elsewhere, as do remnant patches …

Characterizing Cyclin J By Identifying Conserved Protein-Protein Interactions, Phillip Jacob Selman

Wayne State University Theses

Cyclins are proteins that bind to Cyclin-dependent kinases, or Cdks, through a conserved domain called the Cyclin Box. Many Cyclins regulate the cell cycle. A few Cyclins impact cellular processes outside of the cell cycle. Also, a few Cyclins have poorly understood functions.

Cyclin J is a member of the Cyclin superfamily of proteins. Cyclin J is conserved among all metazoans, but is presently not well understood. All the research done on Cyclin J has been done in Drosophila.

Its mRNA is present in the early embryo, then disappears, only to reappear in adult females. When probing protein extracts with …

Regulation Of Nuclear Localization Of The Sole Sumo-Conjugating Enzyme, Ubc9, Palak Sekhri

Wayne State University Theses

The covalent and reversible conjugation of small ubiquitin-like modifier (SUMO) proteins to hundreds of different cellular proteins is catalyzed by a cascade of enzymes including an E1-activating enzyme (SAE1/SAE2), an E2-conjugating enzyme (Ubc9) and multiple E3 ligases. As the only E2 enzyme for SUMO-conjugation, Ubc9 localizes mainly in the nucleus and plays an essential role in regulation of many cellular processes including cell cycle progression through mitosis, cell migration, genome stability, stress response, transcription, and nuclear transport in eukaryotic cells. It is hypothesized that the nuclear localization of Ubc9 is required for efficient sumoylation inside the nucleus because both the …

The Prdm Gene Apoptix Antagonizes Programmed Cell Death In Tribolium, Zahabiya Husain

Wayne State University Theses

Previous studies in holometabolous insects have shown that programmed and cell proliferation play important roles in insect metamorphosis. To elucidate the function of the newly identified Tribolium Prdm gene Apoptix (Apox) I performed a detailed analysis of Apox knockdown effects in the embryo, the pupa and the adult eye. My results revealed that Apox is required for the survival of retinal tissue after onset of differentiation and also generally in tissues which experience high amounts of proliferation and differentiation during pupal development. Further, combinatorial knockdown of Apox and initiator caspases produced evidence that Apox specifically protects from programmed cell death …

Investigation Of A 16s Rna Central Domain Pseudoknot, Jenna Marie Jasinski-Bolak

Wayne State University Theses

X-ray crystallography of the prokaryotic 30S ribosomal subunit revealed a myriad of complex RNA-RNA, RNA-protein, and protein-protein interactions. Among these are several phylogenetically conserved RNA pseudoknots. Pseudoknots are structurally and functionally diverse RNA secondary structures. They are generally formed by two short complimentary sequences separated by many bases of single stranded regions or loops. These relatively simple folds are often yield complex structures that are key components of functionally important conformational changes in RNA structure. One such pseudoknot is located in the central domain of the 16S rRNA.

The central domain pseudoknot is formed by Watson-Crick base pairing between G570-C866 …