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Altering Oligomerization Of Epha2 Via Mutations In The Intracellular Domain, Ryan W. Lingerak 2018 The University of Akron

Altering Oligomerization Of Epha2 Via Mutations In The Intracellular Domain, Ryan W. Lingerak

Honors Research Projects

Eph receptor tyrosine kinases (RTKs) are activated by membrane-bound ligands called ephrins. Eph RTKs are divided into two subclasses, each activated by a specific classes of the ligand ephrin. The overexpression of Eph receptors is correlated to cancer cell metastasis in several different types of cancers. Studies with the EphA2 extracellular domain (ECD) and ephrinA1 ligand have shown that upon binding of ephrin to the receptor, EphA2 undergoes increased oligomerization and activation. This indicates that oligomerization is intimately connected to kinase activity. High resolution crystal structures of the EphA2 ECD have revealed some details of these ligand bound oligomers, as ...


Tea Induction Of Cyp6a8 Promoter In Drosophila Melanogaster, Fiona Retzer 2017 University of Tennessee, Knoxville

Tea Induction Of Cyp6a8 Promoter In Drosophila Melanogaster, Fiona Retzer

University of Tennessee Honors Thesis Projects

No abstract provided.


Regulation Of The Amyloid Precursor Protein By Prostaglandin J2, A Mediator Of Inflammation: Relevance To Alzheimer’S Disease, Teneka L. Jean-Louis 2017 The Graduate Center, City University of New York

Regulation Of The Amyloid Precursor Protein By Prostaglandin J2, A Mediator Of Inflammation: Relevance To Alzheimer’S Disease, Teneka L. Jean-Louis

All Graduate Works by Year: Dissertations, Theses, and Capstone Projects

Inflammation plays a major role in Alzheimer’s disease (AD). Investigating how specific mediators of inflammation contribute to neurodegeneration in AD is crucial. Our studies focused on cyclooxygenases, which are key enzymes in inflammation and highly relevant to AD. Cyclooxygenases (COX -1, constitutive; COX-2, inducible) have emerged as important determinants of AD pathogenesis and progression. COX-2 is highly induced in AD, correlating with AD severity, and COX-1 is also involved in AD. Cyclooxygenases are the rate-limiting enzymes that convert arachidonic acid into prostaglandins (PGs), the principal mediators of CNS neuroinflammation.

The overall GOAL of these studies was to address the ...


Sh2-Dependent Autophosphorylation Within The Tec Family Kinase Itk, Raji E. Joseph, Andrew Severin, Lie Min, D. Bruce Fulton, Amy H. Andreotti 2017 Iowa State University

Sh2-Dependent Autophosphorylation Within The Tec Family Kinase Itk, Raji E. Joseph, Andrew Severin, Lie Min, D. Bruce Fulton, Amy H. Andreotti

Andrew Severin

The Tec family kinase, Itk (interleukin-2 tyrosine kinase), undergoes an in cis autophosphorylation on Y180 within its Src homology 3 (SH3) domain. Autophosphorylation of the Itk SH3 domain by the Itk kinase domain is strictly dependent on the presence of the intervening Src homology 2 (SH2) domain. A direct docking interaction between the Itk kinase and SH2 domains brings the Itk SH3 domain into the active site where Y180 is then phosphorylated. We now identify the residues on the surface of the Itk SH2 domain responsible for substrate docking and show that this SH2 surface mediates autophosphorylation in the full-length ...


The 3-Hydroxyacyl-Acp Dehydratase Component Of The Plant Mitochondrial Fatty Acid Synthase System, Xin Guan, Yozo Okazaki, Andrew Lithio, Ling Li, Xuefeng Zhao, Huanan Jin, Dan Nettleton, Kazuki Saito, Basil J. Nikolau 2017 Iowa State University

The 3-Hydroxyacyl-Acp Dehydratase Component Of The Plant Mitochondrial Fatty Acid Synthase System, Xin Guan, Yozo Okazaki, Andrew Lithio, Ling Li, Xuefeng Zhao, Huanan Jin, Dan Nettleton, Kazuki Saito, Basil J. Nikolau

Ling Li

We report the characterization of the Arabidopsis 3-hydroxyacyl-acyl carrier protein (ACP) dehydratase (mtHD) component of the mitochondrial fatty acid synthase (mtFAS) system, encoded by AT5G60335. The mitochondrial localization and catalytic capability of mtHD were demonstrated with a green fluorescent protein (GFP) transgenesis experiment, and by in vivo complementation and in vitro enzymatic assays. RNAi knockdown lines with reduced mtHD expression exhibit traits typically associated with mtFAS mutants, namely a miniaturized morphological appearance, reduced lipoylation of lipoylated proteins, and altered metabolomes consistent with the reduced catalytic activity of lipoylated enzymes. These alterations are reversed when mthd-rnai mutant plants are grown in ...


Reverse-Genetic Analysis Of The Two Biotin-Containing Subunit Genes Of The Heteromeric Acetyl-Coenzyme A Carboxylase In Arabidopsis Indicates A Unidirectional Functional Redundancy, Xu Li, Hilal Ilarslan, Libuse Brachova, Hui-Rong Qian, Ling Li, Ping Che, Eve Syrkin Wurtele, Basil J. Nikolau 2017 Iowa State University

Reverse-Genetic Analysis Of The Two Biotin-Containing Subunit Genes Of The Heteromeric Acetyl-Coenzyme A Carboxylase In Arabidopsis Indicates A Unidirectional Functional Redundancy, Xu Li, Hilal Ilarslan, Libuse Brachova, Hui-Rong Qian, Ling Li, Ping Che, Eve Syrkin Wurtele, Basil J. Nikolau

Ling Li

The heteromeric acetyl-coenzyme A carboxylase catalyzes the first and committed reaction of de novo fatty acid biosynthesis in plastids. This enzyme is composed of four subunits: biotin carboxyl-carrier protein (BCCP), biotin carboxylase, α-carboxyltransferase, and β-carboxyltransferase. With the exception of BCCP, single-copy genes encode these subunits in Arabidopsis (Arabidopsis thaliana). Reverse-genetic approaches were used to individually investigate the physiological significance of the two paralogous BCCP-coding genes, CAC1A (At5g16390, codes for BCCP1) and CAC1B (At5g15530, codes for BCCP2). Transfer DNA insertional alleles that completely eliminate the accumulation of BCCP2 have no perceptible effect on plant growth, development, and fatty acid accumulation. In ...


Genome Wide Co-Expression Among The Starch Debranching Enzyme Genes Atisa1, Atisa2, And Atisa3 In Arabidopsis Thaliana, Ling Li, Hilal Ilarslan, Martha G. James, Alan M. Myers, Eve Syrkin Wurtele 2017 Iowa State University

Genome Wide Co-Expression Among The Starch Debranching Enzyme Genes Atisa1, Atisa2, And Atisa3 In Arabidopsis Thaliana, Ling Li, Hilal Ilarslan, Martha G. James, Alan M. Myers, Eve Syrkin Wurtele

Ling Li

Each of four starch debranching enzymes (DBE) is distinct and highly conserved across the plant kingdom; however, the specific functions of these proteins in carbohydrate metabolism are not well understood. DBEs function in both biosynthesis and degradation of starch, and two have been shown to function as multimers in various quarternary structures that can contain one or more DBE proteins, i.e. ISA1 homomultimers and ISA1/ISA2 heteromultimers. This study characterizes potential functional relationships between the three isoamylase-type DBE proteins (ISA) of Arabidopsis using a comprehensive bioinformatics analysis and promoter fusion approach to determine tissue-, subcellular-, and temporal specificity of ...


Determining The Role Of Epigenetic Factors In Antifungal Drug Resistance, Abigail R. Gress, Scott D. Briggs, Nina Serratore 2017 Purdue University

Determining The Role Of Epigenetic Factors In Antifungal Drug Resistance, Abigail R. Gress, Scott D. Briggs, Nina Serratore

The Summer Undergraduate Research Fellowship (SURF) Symposium

Epigenetic factors are proteins that regulate gene expression by altering transcriptional machinery access to nucleosomes, DNA wrapped around histone proteins. Two classes of epigenetic factors are ATP-dependent chromatin remodelers and histone modifiers such as histone methyltransferases (HMTs), proteins that add methyl groups to histone tails. This study focuses on AIF4 (Antifungal-Induced Factor 4), a possible HMT induced upon neutral lipid depletion that we hypothesize is regulating antifungal drug resistance genes. Overexpression of AIF4 results in hypersensitivity to antifungal drugs. Studying epigenetic factors in the yeast Saccharomyces cerevisiae, including AIF4, can lead to better understanding of cell adaptation to their environments ...


Tumor Formation In Response To Loss Of Chromatin Remodeler Chd5 In Zebrafish, Taylor R. Sabato, Erin L. Sorlien, Dr. Joseph P. Ogas 2017 Purdue University

Tumor Formation In Response To Loss Of Chromatin Remodeler Chd5 In Zebrafish, Taylor R. Sabato, Erin L. Sorlien, Dr. Joseph P. Ogas

The Summer Undergraduate Research Fellowship (SURF) Symposium

Chromodomain helicase DNA binding protein 5 (CHD5) has been identified as a tumor suppressor in humans. Deletion or mutation of CHD5 has been observed in numerous cancers, including neuroblastoma and melanoma. We hypothesize that chd5 is also a tumor suppressor in zebrafish, a powerful model system to study tumorigenesis. Many genes involved in tumorigenesis are conserved in zebrafish, and they develop fully penetrant tumor phenotypes. We have created chd5 knock-out zebrafish using CRISPR/Cas9 and are monitoring them for tumor development. In addition to the chd5 knock-outs, we are undertaking a double-mutant approach by coupling loss of ...


Another Route For Amino Acid Production?: Reverse Genetic Probing For A Functional Cytosolic Shikimate Pathway In Plants, Gabrielle C. Buck, Joseph Lynch, Natalia Dudareva 2017 Purdue University

Another Route For Amino Acid Production?: Reverse Genetic Probing For A Functional Cytosolic Shikimate Pathway In Plants, Gabrielle C. Buck, Joseph Lynch, Natalia Dudareva

The Summer Undergraduate Research Fellowship (SURF) Symposium

The shikimate pathway is a metabolic pathway that produces the three aromatic amino acids—phenylalanine, tryptophan, and tyrosine—which are essential to human diets and necessary for many plant functions. Consequently, the shikimate pathway is commonly targeted for antibiotic and herbicide strategies as well as genetic engineering in several fields. This pathway is known to be localized in the plastids, or double membrane-bound organelles, of plant cells; however, there is enzymatic evidence of another shikimate pathway in the cell fluid, or cytosol. To determine whether a complete cytosolic shikimate pathway exists, we used a modified gene for the first enzyme ...


Determination Of Amino Acids Involved In Specificity And Activity Of Chladub2, Trent S. Arbough, John M. Hausman, Chittaranjan Das 2017 Purdue University

Determination Of Amino Acids Involved In Specificity And Activity Of Chladub2, Trent S. Arbough, John M. Hausman, Chittaranjan Das

The Summer Undergraduate Research Fellowship (SURF) Symposium

Chlamydia trachomatis is a pathogen which infects humans as a sexually transmitted disease or through ocular infection, causing ocular trachoma. Ocular trachoma is the leading cause of non-congenital blindness in developing countries. The bacteria employs the deubiquitinating enzyme ChlaDUB2 to remove ubiquitin from its inclusion membrane in order to avoid lysosomal degradation. Key amino acids involved in ubiquitin recognition and cleavage were mutated in order to probe substrate specificity and catalytic activity of ChlaDUB2. Mutants were used in fluorometry assays in order to determine how the mutations affect the ability of ChlaDUB2 to release the amino methyl coumarin (AMC) group ...


Determining The Structure Of Phospholipase C Epsilon, Hannah O'Neill, Monita Sieng, Elisabeth Garland-Kuntz, Angeline Lyon 2017 Purdue University

Determining The Structure Of Phospholipase C Epsilon, Hannah O'Neill, Monita Sieng, Elisabeth Garland-Kuntz, Angeline Lyon

The Summer Undergraduate Research Fellowship (SURF) Symposium

The phospholipase C (PLC) epsilon subfamily of PLC enzymes are found at highest concentration within the cardiovascular system. Improper functioning of the enzyme, whether due to overstimulation or changes in expression, has far-reaching effects within the human body Stunted heart valve development and cardiac hypertrophy and are two such examples. The mechanisms by which PLC epsilon activity is regulated in these processes remain unknown, as does the physical structure of the enzyme. In this study, we seek to determine the structure of a PLC epsilon fragment that retains enzymatic activity and is amenable to crystallization. Mutagenesis of PLC epsilon cDNA ...


Performing A Genetic Screen To Identify Factors That Promote Lncrna-Dependent Gene Repression, Chrishan Fernando, Cecilia Yiu, Sara Cloutier, Siwen Wang, Elizabeth Tran 2017 Purdue University

Performing A Genetic Screen To Identify Factors That Promote Lncrna-Dependent Gene Repression, Chrishan Fernando, Cecilia Yiu, Sara Cloutier, Siwen Wang, Elizabeth Tran

The Summer Undergraduate Research Fellowship (SURF) Symposium

Long non-coding RNAs (lncRNAs) were once thought not to have useful functions in organisms but rather to be products of aberrant transcription. However, roles are being found for lncRNAs in beneficial processes such as controlling gene expression. In some of these cases, lncRNAs form R-loops in vivo. R-loops are nucleic acid structures consisting of hybridized strands of single-stranded DNA (ssDNA) and single-stranded RNA (ssRNA) as well as the displaced strand of ssDNA. Formation of these R-loops is important for gene regulation by the lncRNAs. However, factors that promote formation of lncRNA R-loops are not known. The gene PHO84 is being ...


Fluorescent Protein Biosensor For Use In Parkinson's Research, Piper R. Miller, Keelan Trull, Mathew Tantama 2017 Purdue University

Fluorescent Protein Biosensor For Use In Parkinson's Research, Piper R. Miller, Keelan Trull, Mathew Tantama

The Summer Undergraduate Research Fellowship (SURF) Symposium

Purinergic signaling is a type of extracellular communication that occurs between cells, mediated by adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine. In Parkinson’s Disease, purinergic signaling is disrupted, which contributes to neurodegeneration. In order to monitor this change in cell-to-cell signaling, there is a need for the development of a fluorescent protein (FP) biosensor to study the changes in the concentration of the signaling molecule ATP and its decomposition bioproduct ADP. This summer a genetically encoded ADP sensor that measures changes in ADP concentration was developed. This sensor utilizes Forster Resonance Energy Transfer (FRET) which is a sensing ...


Defining The Destruction Box: Understanding How The Apc Recognizes Its Substrates, Hana Maldivita Tambrin, Mark C. Hall 2017 Purdue University

Defining The Destruction Box: Understanding How The Apc Recognizes Its Substrates, Hana Maldivita Tambrin, Mark C. Hall

The Summer Undergraduate Research Fellowship (SURF) Symposium

Cell division is the process by which mother cells give rise to two genetically identical daughter cells. All cells have protein networks to ensure that cell division is completed correctly because mistakes during cell division may cause diseases. The Anaphase-promoting complex (APC) is an important regulatory enzyme that ensures successful completion of mitosis. It acts by removing inhibitors of chromosomal segregation and cytokinesis, as well as other important cell division regulators. Existing chemotherapies, like taxol, act by indirectly inhibiting APC function. This makes APC a potential target for new cancer chemotherapies. However, designing APC inhibitors is challenging because how APC ...


Immunological Strategies To Study Grp170 In Caenorhabditis Elegans, Raven P. Baxter-Christian 2017 State University of New York College at Buffalo - Buffalo State College

Immunological Strategies To Study Grp170 In Caenorhabditis Elegans, Raven P. Baxter-Christian

Biology Theses

Characterization of the ER protein folding chaperone GRP170 of Caenorhabditis elegans could be greatly facilitated by an antibody which recognized the chaperone. Antibodies have not been raised against nematode GRP170. Groups have prepared polyclonal antibodies against vertebrate forms of GRP170. My thesis goal was to investigate whether anti-vertebrate GRP170 antibodies can recognize the nematode homologue on a standard Western Blot assay. Sequences of antigens that groups used to generate anti-vertebrate GRP170 antibodies were analyzed. Peptides used by Ruan et al. (2013), which correspond to regions of human GRP170, shared greatest sequence similarity with nematode GRP170. I investigated whether the Ruan ...


Bacterial Bio-Indicators Of Marcellus Shale Activities In Pennsylvania: A Molecular Ecology Survey, Hephzibah Nwanosike, Jeremy Chen See, Nikea Ulrich, Christopher McLimans, Justin Wright, Maria Campa, Terry Hazen, Jonathan Niles, Regina Lamendella 2017 Juniata College

Bacterial Bio-Indicators Of Marcellus Shale Activities In Pennsylvania: A Molecular Ecology Survey, Hephzibah Nwanosike, Jeremy Chen See, Nikea Ulrich, Christopher Mclimans, Justin Wright, Maria Campa, Terry Hazen, Jonathan Niles, Regina Lamendella

Landmark Conference Summer Research Symposium

The practice of hydraulic fracking has increased over the years especially in Pennsylvania where most of the subterraneous gas-rich Marcellus Shale formations are located. Our previous work showed that headwater streams in proximity to hydraulic fracking operations have significantly different bacterial assemblages as compared to un-impacted streams in central PA. Aquatic bacterial communities are of great importance because they are often the ‘first-responders’ to environmental perturbations. We are interested in which bacteria become enriched, as this might serve as robust biomarkers of fracking, and can potentially biodegrade constituents of fracking fluids. In this study, we plan to expand upon our ...


Expression, Characterization And Metallation Studies Of Human Metallothionein Isoform 2a Using Electrospray Ionization Mass Spectrometry, Devika P. Jayawardena 2017 The University of Western Ontario

Expression, Characterization And Metallation Studies Of Human Metallothionein Isoform 2a Using Electrospray Ionization Mass Spectrometry, Devika P. Jayawardena

Electronic Thesis and Dissertation Repository

Maintenance of the homeostasis of zinc (Zn) is very important in regulating bodily functions. There are over 300 Zn dependent enzymes identified, where Zn plays a structural or catalytic role. However, excess of Zn in a cell is toxic and free Zn ions are tightly controlled. Metallothioneins (MTs) are small cysteine rich proteins, which can bind up to seven Zn ions and act as a Zn reservoir. The MT2a isoform is predominantly found in the liver. My research focused on the overexpression of human MT2a in Escherichia coli and the investigation of Zn binding pathways of MT2a in vitro. At ...


Correction For Sandai Et Al., The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown 2017 University of Aberdeen

Correction For Sandai Et Al., The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown

Janet Walker

No abstract provided.


The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown 2017 University of Aberdeen

The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown

Janet Walker

Microbes must assimilate carbon to grow and colonize their niches. Transcript profiling has suggested that Candida albicans, a major pathogen of humans, regulates its carbon assimilation in an analogous fashion to the model yeast Saccharomyces cerevisiae, repressing metabolic pathways required for the use of alterative nonpreferred carbon sources when sugars are available. However, we show that there is significant dislocation between the proteome and transcriptome in C. albicans. Glucose triggers the degradation of the ICL1 and PCK1 transcripts in C. albicans, yet isocitrate lyase (Icl1) and phosphoenolpyruvate carboxykinase (Pck1) are stable and are retained. Indeed, numerous enzymes required for the ...


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