Genetics and Genomics Commons^™

Epacs: Epigenetic Regulators That Affect Cell Survival In Cancer., Catherine Murari

Theses & Dissertations

Cyclic adenosine monophosphate (cAMP) is a second messenger responsive to many external stimuli, playing an important role in cellular gene expression, metabolism, migration, differentiation, hypertrophy, apoptosis and secretion. All of these cellular functions are important in many diseases including cancer. Most of its effects were initially attributed to the classical protein kinase A (PKA) protein, but cellular functions such as proliferation and migration were found to be PKA independent and dependent on the newly discovered exchange proteins directly activated by cAMP (EPACs). EPACs are single polypeptides that primarily function as guanine exchange factors (GEFs) for Rap proteins that allow the …

Hd2d Is A Regulator Of Abscisic Acid Responses In Arabidopsis, Joshua A. Farhi

Electronic Thesis and Dissertation Repository

Histone deacetylases have important roles in development and stress response in plants. To further investigate their function, the HD2D gene, of the plant specific HD2 family, was studied. An hd2d-1 mutant and two HD2D overexpression lines were used in this study. Germination was delayed in hd2d-1 and HD2D overexpression seeds only in the presence of ABA. HD2D was found to positively regulate the expression of members of the ABA-response pathway (ABI1, ABI5, and RD29A) leading to increased resistance to drought and salinity treatments. Furthermore, HD2D expression delayed flowering by positively regulating FLC expression. Using bimolecular fluorescence complementation, the HD2D protein …

In Vivo Functional Significance Of Ccat2 Long Non-Coding Rna In Myelodysplastic Syndrome, Maitri Y. Shah

Dissertations & Theses (Open Access)

Long non-coding RNAs form the largest part of the mammalian non-coding transcriptome and control gene expression at various levels including chromatin modification, transcriptional and post-transcriptional processing. Although the underlying molecular mechanisms are not yet entirely understood, lncRNAs are implicated in initiation and progression of several cancers. CCAT2 is a lncRNA that spans the highly conserved 8q24 region associated with increased risk for various cancers. CCAT2 has been shown to play an important role in inducing chromosomal instability and supporting cell proliferation and cell cycle arrest. However, a causal role of CCAT2 in initiation of tumorigenesis and the importance of G/T …

Preventing Thymus Involution In K5.Cyclin D1 Transgenic Mice Sustains The Naïve T Cell Compartment With Age, Michelle L. Bolner

Dissertations & Theses (Open Access)

The thymus maintains T cell receptor (TCR) repertoire diversity through perpetual release of self-MHC restricted naive T cells. However, thymus involution during the aging process reduces naïve T cell output, leading to defective immune responsiveness to newly encountered antigens. We have found that early thymus involution precipitates the age-associated shift favoring memory T cell dominancy in young control mice. Furthermore, we have shown that age-related thymus involution is prevented in mice expressing a keratin 5 promoter-driven Cyclin D1 (K5.D1) transgene in thymic epithelial cells (TECs). Thymopoiesis occurs normally in K5.D1 transgenic thymi and sustains T cell output to prevent the …

Rare Occurrences Of Free-Living Bacteria Belonging To Sedimenticola From Subtidal Seagrass Beds Associated With The Lucinid Clam, Stewartia Floridana, Aaron M. Goemann

Masters Theses

Lucinid clams and their sulfur-oxidizing endosymbionts comprise two compartments of a three-stage, biogeochemical relationship among the clams, seagrasses, and microbial communities in marine sediments. A population of the lucinid clam, Stewartia floridana, was sampled from a subtidal seagrass bed at Bokeelia Island Seaport in Florida to test the hypotheses: (1) S. floridana, like other lucinids, are more abundant in seagrass beds than bare sediments; (2) S. floridana gill microbiomes are dominated by one bacterial operational taxonomic unit (OTU) at a sequence similarity threshold level of 97% (a common cutoff for species level taxonomy) from 16S rRNA genes; …

A Unified Framework For The Prioritization Of Variants Of Uncertain Significance In Hereditary Breast And Ovarian Cancer Patients, Natasha G. Caminsky

Electronic Thesis and Dissertation Repository

A significant proportion of hereditary breast and ovarian cancer (HBOC) patients receive uninformative genetic testing results, an issue exacerbated by the overwhelming quantity of variants of uncertain significance identified. This thesis describes a framework where, aside from protein coding changes, information theory (IT)-based sequence analysis identifies and prioritizes pathogenic variants occurring within sequence elements predicted to be recognized by proteins involved in mRNA splicing, transcription, and untranslated region binding and structure. To support the utilization of IT analysis, we established IT-based variant interpretation accuracy by performing a comprehensive review of mutations altering mRNA splicing in rare and common diseases.

Custom …

Characterization Of The Atsr/Atst Global Regulatory Pathway In Burkholderia Ceocepacia, Maryam Khodai-Kalaki

Electronic Thesis and Dissertation Repository

Phosphorylation cascades governed by two-component signal transduction systems provide key signalling mechanisms in bacteria, simple eukaryotes and higher plants, allowing them to translate signals into adaptive responses. These regulatory pathways consist of a transmembrane sensor protein that responds to an environmental cue leading to autophosphorylation, followed by the transfer of the phosphate to a cytoplasmic response regulator. Here, I study AtsR, a membrane-bound hybrid sensor kinase of Burkholderia cenocepacia, that negatively regulates quorum sensing related virulence factors such as biofilm, type 6-secretion and protease secretion. B. cenocepacia is a Gram-negative opportunistic pathogen which causes severe, chronic respiratory infections in …

Genetic Analysis Of A Non-Germinating Mutant Of Arabidopsis Thaliana, Md Jakir Hossan

Electronic Thesis and Dissertation Repository

Seed germination is partially controlled by plant hormone gibberellins (GAs). Chemical mutagenesis yielded an Arabidopsis thaliana mutant gm11, which has an absolute gibberellin requirement for seed germination. This mutant exhibited phenotypes of GA-rescuable dwarfs, including dark-green leaves, and reduced fertility. However, with repeated GA treatment, gm11 develops into fertile plants with a nearly wild type phenotype. Bulked-segregant analysis mapped gm11 to the bottom arm of chromosome 1, and subsequent next-generation mapping revealed that the mutation is a G → A transition in At1g79460 (GA2), creating a premature stop codon. This gene encodes an ent-kaurene synthase (KS) which catalyzes …

Determination Of The Phylogenetic Range Of Sex Combs Reduced Activity In Drosophila Melanogaster, Laura E. Garofalo

Electronic Thesis and Dissertation Repository

The homeotic selector (Hox) genes are required for body patterning in bilaterians. Sex combs reduced (SCR) is a HOX protein in Drosophila melanogaster with two activities: SCR^T1 and SCR^lab activity required for patterning the prothorax (T1) and labia, respectively. SCR^T1 is proposed to be conserved throughout bilaterians while the phylogenetic range of functional conservation of SCR^lab is comparatively unknown. The goal of this work was to elucidate the evolutionary time point at which SCR activity changed. CRISPR/Cas9 transgenesis was used to incorporate ɸC31 integrase recombination sites in Drosophila Scr. The ɸC31 integrase could …

Calmodulin-Like Protein 38: A Component Of Ribonucleoprotein Particles During Hypoxic Stress Responses In Arabidopsis, Ansul Lokdarshi

Doctoral Dissertations

Waterlogging stress leads to a crisis in energy metabolism and the accumulation of toxic metabolites due to the hypoxic and/or anoxic environment associated with this condition. To respond and adapt to this situation, higher plants employ an integrated genetic program that leads to the induction of anaerobic response polypeptide genes that encode metabolic and signaling proteins involved in altering metabolic flow and other adaptive responses. The study presented here shows that the Arabidopsis thaliana calmodulin-like protein CML38 is calcium sensor protein that serves as a member of the core anaerobic response gene family and is involved in modulating the survival …

Identification Of Familial Wilms Tumor Predisposition Genes Using Whole Genome Sequencing, Timothy B. Palculict

Dissertations & Theses (Open Access)

Wilms tumor, a childhood tumor arising from undifferentiated renal mesenchyme, is diagnosed in North America at a frequency of 1 in 10,000 live births and accounts for 5% of all pediatric cancers. The etiology of Wilms tumor is heterogeneous with multiple genes known to have an effect on Wilms tumor development; however, these genes are rarely associated with familial Wilms tumor. Gene mutations in WT1, WTX, CTNNB1 and TP53 are observed in a third of sporadic tumors, while the causative gene(s) responsible for familial Wilms tumor are largely unknown. Approximately 2% of Wilms tumor patients have a family …

Hippocalcin Response To Calcium: Do Conserved Tryptophans – W30 Or W103 – Matter?, Sunkesula K. Sagar

Graduate School of Biomedical Sciences Theses and Dissertations

Changes in intracellular calcium levels play a very important role in cell signaling, in turn, affecting neuronal functions such as memory, learning and cell death. A class of proteins called Neuronal Calcium Sensor (NCS) proteins serves to modulate the functioning of the neuronal cells in response to changes in calcium levels, and prevent neuronal apoptosis. Structurally, all NCS proteins have 4 calcium-binding EF hand motifs, although EF1 does not bind to calcium in many members. All NCS proteins have an acyl modification at the N- terminus – where a myristoyl group is added post-translationally. Hippocalcin (HPCA) is an NCS protein, …

Functional Analysis Of Synthetic Gene Circuits Controlling A Protein Pump In Yeast, Junchen Diao

Dissertations & Theses (Open Access)

Synthetic biology aims to build biological devices to understand living systems and explore new applications. Synthetic gene circuits such as genetic switches, oscillators and logic gates are at the core of many synthetic biology applications. These gene circuits often include a sensor/regulator protein capable to detect small molecules and then transduce them into a regulatory signal to generate measurable output. Similar signal transduction networks are also abundant in nature. However, in many natural and engineered scenarios, the output also affects the regulator/sensor protein. How such interactions between the regulator/sensor and the output affect synthetic gene circuit function has not been …

Distinct Physiological Roles For The Two Isoforms Of The Er Chaperone Grp170 In Caenorhabditis Elegans, Yuanyuan Li

Biology Theses

GRP170 is a large molecular chaperone found in the ER of all eukaryotes. The nematode Caenorhabditis elegans has two loci encoding GRP170: T24H7.2 (grp170a) and T14G8.3 (grp170b). The phenotypes of nematodes genetically deficient for either grp170a or grp170b were compared to a standard laboratory strain with functional grp170 loci. Worms that were deficient for grp170a developed 32% slower than the control strain. The loss of grp170a had a significant but modest reduction on the life span compared to the control strain. Worms deficient for grp170a also displayed significantly increased embryonic lethality and resulted in 6.9% arrested embryos. The loss of …

Assessment Of Genome-Wide Genetic And Epigenetic De Novo Variation In Families With Monozygotic Twins Discordant For Schizophrenia, Christina A. Castellani

Electronic Thesis and Dissertation Repository

Schizophrenia (OMIM: 181500) is a common, debilitating and life-altering disorder. It affects 1% of the population worldwide and most often presents in early adulthood leading to devastating effects for patients, their families and society. Despite thousands of studies performed on the underlying mechanisms of schizophrenia, the causes of the disease remain unknown. However, what is known is that environmental, genetic and epigenetic factors contribute to the development of this complex disorder. Although a genetic role in schizophrenia is well established, the search for schizophrenia genes using traditional approaches has remained challenging. Interestingly, monozygotic twins show concordance for schizophrenia only 50% …

The Effects Of Sustained, High-Velocity Exercise On Gene Expression In California Yellowtail (Seriola Lalandi), Kelli Hatter

Undergraduate Honors Theses

California Yellowtail muscle fibers have been observed to exhibit two drastically different development patterns resulting from the speeds at which they are exercised. When fish are exercised at a moderate rate their epaxial fast-twitch muscle fibers grow in diameter—hypertrophy; when they are exercised at a fast speed, more new epaxial fast-twitch muscle fibers are produced—hyperplasia. To determine the underlying reason for this difference in muscle development, my summer research project and honors thesis exercised fish at: fast, moderate, and control speeds for a sustained amount of time to determine what is happening on a cellular level to cause the observed …

Structural And Functional Interactions Between Bro1 Domain Of Human Alix Protein And Nucleocapsid Packaging Rna Complex From Hiv, Scott Gross

Graduate School of Biomedical Sciences Theses and Dissertations

A virus is only as powerful as its ability to spread. Enveloped retroviruses, namely HIV-1, use exocytosis pathways that normal host cells use to release particles from the plasma membrane. The main pathways of interest in this study are the Endosomal Sorting Complex Required for Transport (ESCRT) and adjacent ALIX pathways. The ESCRT pathway is especially important for degradation of receptor/cargo complexes that form Multi-Vesicular Bodies (MVBs). Currently, there is no known therapy that targets this endosomal pathway, which would prevent the spread of the virus to other cells. The virus has adapted to jump from pathway to pathway when …

A Phylogenetic Analysis Of Dictyostelium Purpureum Based On Nuclear Rdna Sequences, Mahmoud Suliman

Graduate Theses and Dissertations

Dictyostelids (cellular slime molds) are eukaryotic microorganisms that have both unicellular and multicellular stages during their life cycle. In this study, a molecular phylogenetic analysis was conducted for isolates of one species (Dictyostelium purpureum) based DNA sequences of the ITS, 5.8S and SSU regions of nuclear rDNA. Moreover, a detailed morphological study was carried out using images obtained with both dissecting and compound microscopes. Mating experiments were carried out to assess macrocysts formation between each pair of isolates. The constructed molecular phylogenetic trees indicate that (1) D. purpureum isolates are more closely related to each other than to other species …

Characterization Of Putative Exbb And Exbd Leads To The Identification Of A Potential Tol-Pal System In Rhizobium Leguminosarum Atcc 14479, Valeria Barisic

Electronic Theses and Dissertations

Rhizobium leguminosarum is a Gram negative nitrogen-fixing soil bacterium. Due to the limited bioavailability of iron, bacteria utilize siderophores that scavenge and bind available iron. The transport of iron-siderophore complexes is achieved by the TonB-ExbB-ExbD complex. We have previously shown that a functional TonB protein is necessary for iron transport by creating ΔtonB mutants and assessing their growth and 55Fe-siderophore transport ability. We attempted to identify and characterize the roles of putative exbB and exbD genes using a similar approach. Growth curves and sequence analyses suggest putative exbB and exbD may be the tolpal-associated genes tolQ and tolR. Phenotypic and …

Determination Of The Dermacentor Variabilis Ferritin Gene Exon And Intron Structure, Dana Sylvestre

Honors College Theses

The American dog tick Dermacentor variabilis is a hard body, 3-host tick that relies on the blood of small and large mammals to satisfy its metabolic demands. The excess iron consumed in its blood diet can cause cellular damage through oxidation. Ticks rely on the ferritin protein to metabolize iron. For this reason, it is important to further characterize the ferritin gene. After extracting DNA from D. variabilis ticks, the suspected ferritin gene was amplified through polymerase chain reaction (PCR) and purified. The putative ferritin DNA fragment was inserted into plasmids and isolated in a long PCR length gene cloning. …

Dna Polymerase Θ (Polq) And The Cellular Defense Against Dna Damage, Matthew J. Yousefzadeh

Dissertations & Theses (Open Access)

In mammalian cells, DNA polymerase θ (POLQ) is an unusual specialized DNA polymerase whose in vivo function is under active investigation. The protein is comprised of an N-terminal helicase-like domain, a C-terminal DNA polymerase domain, and a large central domain that spans between the two. This arrangement is also found in the Drosophila Mus308 protein, which helps confer resistance to DNA interstrand crosslinking agents. Homologs of POLQ and Mus308 are found in eukaryotes, including plants, but a comparison of phenotypes suggests that not all of these genes are functional orthologs. Flies with defective Mus308 are sensitive to DNA interstrand crosslinking …

Characterization Of A Novel Clade Of Transporters In Phytophthora, Stephanie Padula, Paul F. Morris Dr, Howard Casey Cromwell Dr., Menaka Ariyaratne, Andrew Wagner

Honors Projects

The oomycete Phytophthora parasitica has a worldwide distribution and is an economically important pathogen of more than 100 species⁴. RNA-seq analysis showed that one gene, PPTG_16698 has the 5^th highest level of expression of all transport proteins in the zoospore stage, and is highly conserved throughout Phytophthora species. This project attempts to characterize the important biological role that PPTG_16698 plays in P. parasitica and other oomycetes. Three strategies have been implemented to accomplish this goal: growth analysis by heterologous expression in yeast, metabolite analysis in yeast, and construction of a GFP fusion protein to enable localization of …

Investigating The Roles Of P63 And P73 Isoforms To Therapeutically Treat P53-Altered Cancers, Avinashnarayan Venkatanarayan

Dissertations & Theses (Open Access)

Investigating the roles of p63 & p73 isoforms to therapeutically treat

p53-altered cancers

Avinashnarayan Venkatanarayan, M.S.

Supervisory Professor: Elsa R. Flores, Ph.D.

The TP53 tumor suppressor is mutated in approximately 50% of human cancers rendering cancer therapies ineffective. p53 reactivation suppresses tumor formation in mice. However, this strategy has proven difficult to implement therapeutically. An alternate approach to overcome p53 loss is to manipulate the p53-family members, p63 and p73, which interact and share structural similarities to p53. p63 and p73, unlike p53 are less frequently mutated and have two major isoforms with distinct functions …

Tracking Plastid Gene Migration In Karenia Brevis, Kelly E. Scott

Honors Theses

Karenia brevis is a marine dinoflagellate responsible for the harmful algal blooms (also known as red tides) in the Gulf of Mexico. K. brevis expresses antisense (AS) RNAs, each of which has a complementary region to the messenger RNA (mRNA) of a variety of genes. In dinoflagellates, many plastid (and mitochondrial) genes have migrated to the nuclear genome. It is unknown whether chloroplast genes, such as photosystem – D2, have migrated in K. brevis. It is also unknown where the gene that expresses the AS RNA for photosystem D2 resides. The protein-coding gene and the AS RNA-expressing gene could …

Characterization Of Head Involution Defective (Hid) As A Pro-Apoptotic Gene In Megasalia Scalaris, Hakyong Kwak

Chancellor’s Honors Program Projects

No abstract provided.

Mir156 Participates In Iron Homeostasis Through Targeting Spl9/Spl15 In Arabidopsis Thaliana, Ying Wang

Electronic Thesis and Dissertation Repository

Plants growing under iron deficiency suffer from multiple physiological defects. Although the effects of microRNA156 (miR156) on multiple aspects of plant development have been investigated, a possible role of miR156 in plant iron homeostasis has not been shown. By employing next-generation RNA-sequencing, the current research demonstrated that multiple iron homeostasis-related genes, including ones coding for Ferritins, group Ib bHLH transcription factors, and key enzymes involved in iron uptake, were differentially expressed in Arabidopsis thaliana plants overexpressing miR156. Overexpression of miR156 also enhanced Arabidopsis growth under iron-deficiency. In addition, expression analysis revealed that miR156 is a positive regulator of FIT and …

Relative Abundances Of The Recently Introduced Barnacles, Megabalanus Coccopoma And An Unidentified Species Of Megabalanus, In The Southeastern U.S., Jennifer L. Tyson

Honors College Theses

ABSTRACT

Megabalanus coccopoma is a prominent invasive species off the coast of Georgia. Recently, among collected samples thought to be M. coccopoma, several individuals of an unidentified species of barnacle were found. The species has been identified as a Megabalanus species, but is still unidentified to the species level. Species identification is difficult due to morphological variation, inconsistent taxonomic keys, and unknown origin. In this study I developed a method to accurately distinguish M. coccopoma from the unidentified Megabalanus sp. using sequence differences in the mitochondrial Cytochrome Oxidase I (COI) gene. This study will provide an accurate estimate of …

The E26 Transformation-Specific Transcription Factors Pu.1, Spi-B, And Spi-C Regulate Transcriptional Activation And Repression Of Nfkb1 To Control B Cell Development And Function, Stephen Ka Ho Li

Electronic Thesis and Dissertation Repository

PU.1, Spi-B, and Spi-C are highly related E26 transformation-specific family transcription factors that can bind nearly identical DNA sequences. PU.1 and Spi-B (encoded by Spi1 and Spib respectively) are important for B cell development and function, but the function of Spi-C (encoded by Spic) in B cells is not clear. The objective of this study was to determine PU.1, Spi-B, and Spi-C’s function during B cell development, and during TLR-mediated responses. It was hypothesized that PU.1 and Spi-B were required for positively regulating components of TLR responses, and Spi-C inhibited PU.1 and Spi-B targets. Spi1^+/-Spib^-/- ( …

The Effect Of Serrate Transmembrane Domain Substitution On Notch Signaling, James Z. Curlin

Senior Theses and Projects

The Notch signaling pathway is a crucial means by which organisms differentiate cells during development. Notch is regulated primarily through the interaction of a Notch receptor protein and a ligand protein, in two specific ways. Cis-inhibition occurs when both a ligand and receptor are present on the same cellular membrane. This results in the cis-ligand binding to the receptor and preventing the ligand on an adjacent cell from binding and activating the receptor. Alternatively, trans-activation occurs when the ligand and receptor are on adjacent cells, and results in the activation of the Notch pathway. Both the receptor …

Soybean Isoflavonoid Biosynthesis: Constituents And Circumstance At The Transcriptomic And Molecular Levels, Mehran Dastmalchi

Electronic Thesis and Dissertation Repository

Isoflavonoids are specialized metabolites, almost exclusive to the legume family of plants. They are actors in symbiosis with nitrogen-fixing bacteria and in plant stress response. Isoflavonoids are noted for their human health benefits. Isoflavonoid content in legumes has proven to be a complex trait. The goal of the present research is to determine the mechanisms underlying isoflavonoid biosynthesis in soybean.

The first approach was to unravel the genetic factors of isoflavonoid biosynthesis. A branch-point enzyme of the phenylpropanoid pathway, chalcone isomerase (CHI), catalyzes the reaction producing flavanones, the nucleus for many downstream metabolites such as isoflavonoids. I identified twelve soybean …