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Multivalency Regulates Activity In An Intrinsically Disordered Transcription Factor, Sarah Clark, Janette B. Myers, Ashleigh King, Radovan Fiala, Jiri Novacek, Grant Pearce, Steve L. Reichow, Elisar J. Barbar 2018 Department of Biochemistry and Biophysics, Oregon State University

Multivalency Regulates Activity In An Intrinsically Disordered Transcription Factor, Sarah Clark, Janette B. Myers, Ashleigh King, Radovan Fiala, Jiri Novacek, Grant Pearce, Steve L. Reichow, Elisar J. Barbar

Chemistry Faculty Publications and Presentations

The transcription factor ASCIZ (ATMIN, ZNF822) has an unusually high number of recognition motifs for the product of its main target gene, the hub protein LC8 (DYNLL1). Using a combination of biophysical methods, structural analysis by NMR and electron microscopy, and cellular transcription assays, we developed a model that proposes a concerted role of intrinsic disorder and multiple LC8 binding events in regulating LC8 transcription. We demonstrate that the long intrinsically disordered C-terminal domain of ASCIZ binds LC8 to form a dynamic ensemble of complexes with a gradient of transcriptional activity that is inversely proportional to LC8 occupancy. The preference ...


Development Of Endogenous Tagging Plasmids For Characterization Of Protein Interactions, Localization, And Post-Translational Modifications Of Tetrahymena Thermophila Rad23, Evan Andrew Wilson 2018 Missouri State University

Development Of Endogenous Tagging Plasmids For Characterization Of Protein Interactions, Localization, And Post-Translational Modifications Of Tetrahymena Thermophila Rad23, Evan Andrew Wilson

MSU Graduate Theses

Rad23 is a protein involved in both nucleotide excision repair (NER) and proteasome-mediated degradation, and has been suggested to facilitate interactions between these two pathways. The model organism Tetrahymena thermophila, which has a transcriptionally silent micronucleus, provides a useful platform for studying the role of Rad23 in global genome NER (GG-NER). However, the ectopic expression systems used thus far in T. thermophila to study Rad23 are repressed by UV light and do not account for the background expression of endogenous RAD23; these phenomena prevent insightful gains to the true dynamics of Rad23. In this thesis, endogenous tagging cassettes have been ...


The Regulation Of Dna Methylation In Mammalian Development And Cancer, Nicolas Veland 2018 The University of Texas M D Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

The Regulation Of Dna Methylation In Mammalian Development And Cancer, Nicolas Veland

UT GSBS Dissertations and Theses (Open Access)

DNA methylation is an essential epigenetic modification in mammals, as it plays important regulatory roles in multiple biological processes, such as gene transcription, maintenance of chromosomal structure and genomic stability, genomic imprinting, retrotransposon silencing, and X-chromosome inactivation. Dysregulation of DNA methylation is associated with various human diseases. For example, cancer cells usually show global hypomethylation and regional hypermenthylation, which have been implicated in genomic instability and tumor suppressor silencing, respectively. Although great progress has been made in elucidating the biological functions of DNA methylation over the last several decades, how DNA methylation patterns and levels are regulated and dysregulated is ...


Investigating The Impact Of Intragenic Dna Methylation On Gene Expression, And The Clinical Implications On Tumor Cells And Associated Stroma, Michael McGuire 2018 The University of Texas M D Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

Investigating The Impact Of Intragenic Dna Methylation On Gene Expression, And The Clinical Implications On Tumor Cells And Associated Stroma, Michael Mcguire

UT GSBS Dissertations and Theses (Open Access)

Investigations into the function of non-promoter DNA methylation have yielded new insights into epigenetic regulation of gene expression. Previous studies have highlighted the importance of distinguishing between DNA methylation in discrete functional regions; however, integrated non-promoter DNA methylation and gene expression analyses across a wide number of tumor types and corresponding normal tissues have not been performed. Through integrated analysis of gene expression and DNA methylation profiles, we uncovered an enrichment of DNA methylation sites within the gene body and 3’UTR in which DNA methylation is strongly positively correlated with gene expression. We examined 32 tumor types and identified ...


Functional Similarity Of Prd-Containing Virulence Regulators In Bacillus Anthracis, Malik Raynor 2018 The University of Texas M D Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

Functional Similarity Of Prd-Containing Virulence Regulators In Bacillus Anthracis, Malik Raynor

UT GSBS Dissertations and Theses (Open Access)

Bacillus anthracis produces three regulators, AtxA, AcpA, and AcpB, that control virulence gene expression and are members of an emerging class of regulators termed “PCVRs” (Phosphoenolpyruvate-dependent phosphotransferase regulation Domain-Containing Virulence Regulators). AtxA controls expression of the toxin genes; lef, cya, and pag, and is the master virulence regulator and archetype PCVR. AcpA and AcpB are less well studied. AcpA and AcpB independently positively control transcription of the capsule biosynthetic operon capBCADE, and culture conditions that enhance AtxA activity result in capBCADE transcription in strains lacking acpA and acpB. RNA-Seq was used to assess the regulons of the paralogs in strains ...


Identification And Utility Of Dna In Exosomes, Paul Kurywchak 2018 The University of Texas M D Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

Identification And Utility Of Dna In Exosomes, Paul Kurywchak

UT GSBS Dissertations and Theses (Open Access)

Cancer-associated mortality has been declining for two decades but remains a significant public health problem, especially when patients initially present with advanced disease. Early detection methods have improved survival rates but remain unavailable for a majority of cancers due to a lack of sensitive biomarkers or numerous limitations associated with current diagnosis strategies. Approaches to develop “liquid biopsies” by detecting tumor cells or DNA in the blood have led to several breakthroughs and create the potential for non-invasive, routine assessment of diseases status. However, these biomarkers are rare and currently difficult to isolate, especially in the early stages of disease ...


Trim24 In Normal & Malignant Hematopoiesis, Justin Shaw 2018 The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

Trim24 In Normal & Malignant Hematopoiesis, Justin Shaw

UT GSBS Dissertations and Theses (Open Access)

Treatment for acute myeloid leukemia (AML) has changed little in the past four decades. For the majority of AML patients, current treatment options include chemotherapy and allogeneic stem cell transplants, which also involves high-dose chemotherapy or radiation treatment. These options have little success in the long-run, as only an estimated 26% of patients survive five years post-diagnosis. In efforts to address this low survival rate, interest has increased for targeting epigenetic pathways in AML. This focus stems from the discovery that AML is frequently driven by blockades on hematopoietic stem cell differentiation, which involves a series of coordinated epigenetic changes ...


Phosphorylation Impairs Dicer1 Function To Accelerate Aging And Tumorigenesis In Vivo, Neeraj Aryal 2018 The University of Texas M D Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences

Phosphorylation Impairs Dicer1 Function To Accelerate Aging And Tumorigenesis In Vivo, Neeraj Aryal

UT GSBS Dissertations and Theses (Open Access)

Altered DICER1 protein levels are associated with developmental disorders, infertility, macular degenerative blindness, aging, and cancer in humans. Recently, post-translational regulation of Dicer1 via phosphorylation has been described in C. elegans. Oscillation of Dicer1 phosphorylation to regulate its activity is essential for germ cell development and embryogenesis in worms. These observations led us to posit that Dicer1 protein levels and activity are under tight regulation for normal mammalian homeostasis. To test whether phosphorylation of Dicer1 regulates its activity in mammals, I generated phospho-mimetic knock-in mouse models by replacing Serines 1712 and 1836 with Aspartic acids individually or together (dual phosphorylation ...


Investigation For Novel Anti-Apoptotic Factors In The Neurons Of Drosophila Melanogaster, Haylie Rachel Lam 2018 University of Tennessee, Knoxville

Investigation For Novel Anti-Apoptotic Factors In The Neurons Of Drosophila Melanogaster, Haylie Rachel Lam

Chancellor’s Honors Program Projects

No abstract provided.


Involvement Of The Ino80 Complex In Chromosome Segregation, Jesus Moreno 2018 University of Arkansas, Fayetteville

Involvement Of The Ino80 Complex In Chromosome Segregation, Jesus Moreno

Theses and Dissertations

Chromatin remodeling complexes are multi-protein complexes that regulate the dynamics of the nucleosomes in the genome. The INO80 chromatin remodeling complex participates in varied biological processes such as: transcription, DNA repair, DNA replication and chromosome integrity. It catalyzes the eviction of the H2A.Z variant histone as well as whole nucleosome eviction. This complex is comprised of 15 subunits and the contribution of each to chromosome segregations remains unknown. To evaluate the contribution of each subunit to chromosome segregation, we tested deletion mutants of the non-essential subunits for DNA content and benomyl sensitivity. Also, we assessed members of the SWR1 ...


Unravelling The Layers Of Cell Wall Synthesis And Function In Rice, Ritu Mihani 2018 University of Arkansas, Fayetteville

Unravelling The Layers Of Cell Wall Synthesis And Function In Rice, Ritu Mihani

Theses and Dissertations

The plant cell wall is of critical importance to plant growth and survival, functioning in maintaining structural integrity, supporting cell expansion, and acting as the first line of defense in response to biotic and abiotic stresses. The major components of the cell wall are cellulose, hemicelluloses, lignin, and pectin. Recent focus on the transcriptional machinery regulating cell wall biosynthesis in plants has revealed many key transcription factors responsible for orchestrating cell wall deposition. However, many of these TFs act redundantly and work coherently with a suite of TFs to activate the cell wall biosynthetic machinery. Heterologous expression of TFs is ...


Genotype-Specific Insertion Of Cytotoxic Genetic Elements Into Cancer Cells, Ryan Englander 2018 University of Connecticut

Genotype-Specific Insertion Of Cytotoxic Genetic Elements Into Cancer Cells, Ryan Englander

University Scholar Projects

The new gene editing system CRISPR/Cas9, composed of a complex composed of a guide RNA and the Cas9 endonuclease, promises to revolutionize biological research and potentially allow clinicians to directly modify patient DNA in vivo. While its applications in the treatment of genetic diseases and in modifying immune cells for immunotherapy are currently being explored, CRISPR/Cas9’s potential utility as a modular system for targeting tumor-specific mutated sequences has not as of yet been explored. While CRISPR/Cas9 is specific enough to target small insertions and deletions or gross chromosomal rearrangements, it is not specific enough to reliably ...


Neutrophils From Both Susceptible And Resistant Mice Efficiently Kill Opsonized Listeria Monocytogenes, Michelle G. Pitts, Travis A. Combs, Sarah E. F. D'Orazio 2018 University of Kentucky

Neutrophils From Both Susceptible And Resistant Mice Efficiently Kill Opsonized Listeria Monocytogenes, Michelle G. Pitts, Travis A. Combs, Sarah E. F. D'Orazio

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Inbred mouse strains differ in their susceptibility to infection with the facultative intracellular bacterium Listeria monocytogenes, largely due to delayed or deficient innate immune responses. Previous antibody depletion studies suggested that neutrophils (polymorphonuclear leukocytes [PMN]) were particularly important for clearance in the liver, but the ability of PMN from susceptible and resistant mice to directly kill L. monocytogenes has not been examined. In this study, we showed that PMN infiltrated the livers of BALB/c/By/J (BALB/c) and C57BL/6 (B6) mice in similar numbers and that both cell types readily migrated toward leukotriene B4 in an in ...


The Characterization Of The Transcription Factor Msab And Its Role In Staphylococcal Virulence, Justin Batte 2018 The University of Southern Mississippi

The Characterization Of The Transcription Factor Msab And Its Role In Staphylococcal Virulence, Justin Batte

Dissertations

Staphylococcus aureus is a common human pathogen that is responsible for a wide range of infections, ranging from relative minor skin infections to life-threatening disease such as bacteremia, septicemia, and endocarditis. S. aureus possesses many different virulent factors that aid in its ability to cause this wide array of infections. One major virulence factor includes the production of capsular polysaccharide (CP). The production of CP plays a major role in the virulence response during infection specifically by providing S. aureus an antiphagocytic mechanism that allows the pathogen to evade phagocytosis during an infection. S. aureus has developed complex genetic regulatory ...


Investigating The Role Of Dnc-2 And Dli-1 On Ampa Receptor Mediated Behaviors In Caenorhabditis Elegans, Natalie Oberding 2018 Carroll College, Helena, MT

Investigating The Role Of Dnc-2 And Dli-1 On Ampa Receptor Mediated Behaviors In Caenorhabditis Elegans, Natalie Oberding

Life and Environmental Sciences Undergraduate Theses

Γ-amino-3-hydroxy-5methylisoxazole-4-proprionic acid (AMPA) receptors (AMPARs) are protein complexes involved in excitatory neurotransmission. AMPARs are tetrameric structures consisting of the pairings of GluA1, GluA2, GluA3, and GluA4 subunits. Dctn2 and Dync1li1 were found to interact with GluA2 in an immunoprecipitation screen that was performed on postnatal day 14 rat brains. DNC-2 and DLI-1 were identified as the C. elegans homologs of Dctn2 and Dync1li1, respectively, and are components of motor proteins that transport cargo throughout the cell. The goal of this project was to determine whether knocking down DNC-2 and DLI-1 by RNAi affects AMPA Receptor-mediated behaviors, including mechanosensation and chemosensation ...


A Genetic Screen To Identify Fission Yeast Genes With Roles In Protecting Against Perturbation Of The Actin Cytoskeleton, Dorota Michalski 2018 The University of Western Ontario

A Genetic Screen To Identify Fission Yeast Genes With Roles In Protecting Against Perturbation Of The Actin Cytoskeleton, Dorota Michalski

Electronic Thesis and Dissertation Repository

In the fission yeast, Schizosaccharomyces pombe, stress upon the cell division machinery leads to the activation of a cytokinesis checkpoint. This checkpoint results in a delay in cell cycle progression and the prolonged maintenance of a cytokinesis competent cellular state. In this state the cell is able to continuously reform/repair the actomyosin ring until cell division is achieved. To uncover genes that play a role in enforcing this checkpoint, the actin depolymerizing drug Latrunculin A (LatA) was used to perturb the cytokinetic machinery in a set of 3400 viable haploid S. pombe gene deletion mutants. Thirty-eight gene deletion mutants ...


Oligonucleotides Targeting Tcf4 Triplet Repeat Expansion Inhibit Rna Foci And Mis-Splicing In Fuchs' Dystrophy, Jiaxin Hu, Ziye Rong, Xin Gong, Zhengyang Zhou, Vivek K. Sharma, Chao Xing, Jonathan K. Watts, David R. Corey, V. Vinod Mootha 2018 UT Southwestern Medical Center

Oligonucleotides Targeting Tcf4 Triplet Repeat Expansion Inhibit Rna Foci And Mis-Splicing In Fuchs' Dystrophy, Jiaxin Hu, Ziye Rong, Xin Gong, Zhengyang Zhou, Vivek K. Sharma, Chao Xing, Jonathan K. Watts, David R. Corey, V. Vinod Mootha

RNA Therapeutics Institute Publications

Fuchs' endothelial corneal dystrophy (FECD) is the most common repeat expansion disorder. FECD impacts 4% of U.S. population and is the leading indication for corneal transplantation. Most cases are caused by an expanded intronic CUG tract in the TCF4 gene that forms nuclear foci, sequesters splicing factors and impairs splicing. We investigated the sense and antisense RNA landscape at the FECD gene and find that the sense-expanded repeat transcript is the predominant species in patient corneas. In patient tissue, sense foci number were negatively correlated with age and showed no correlation with sex. Each endothelial cell has approximately 2 ...


Rna Interference Of Mdr Genes In Leptinotarsa Decemlineata And Trichoplusia Ni By Ingestion Of Double-Stranded Rna, Grant Favell 2018 The University of Western Ontario

Rna Interference Of Mdr Genes In Leptinotarsa Decemlineata And Trichoplusia Ni By Ingestion Of Double-Stranded Rna, Grant Favell

Electronic Thesis and Dissertation Repository

Insecticide resistance is often linked to the expression of detoxification genes such as MDRs which code for ATP-binding cassette efflux transporters with broad substrate specificity. To investigate the role of MDR genes in Leptinotarsa decemlineata and Trichoplusia ni tolerance for the insecticide ivermectin, ingested dsRNA was used to attempt silencing of various MDR genes in these insects through RNA interference. Silencing was effective in L. decemlineata, but not T. ni. No change in ivermectin tolerance was found in L. decemlineata after MDR gene silencing. Because RNAi efficiency was different between L. decemlineata and T. ni, the stability of dsRNA in ...


Copy Number Variation In The Porcine Genome Detected From Whole-Genome Sequence, Rebecca Anderson 2018 University of Nebraska-Lincoln

Copy Number Variation In The Porcine Genome Detected From Whole-Genome Sequence, Rebecca Anderson

Honors Theses, University of Nebraska-Lincoln

Copy number variations (CNVs) are large insertions, deletions, and duplications in the genome that vary between individuals in a species. These variations are known to impact a broad range of phenotypes from molecular-level traits to higher-order clinical phenotypes. CNVs have been linked to complex traits in humans such as autism, attention deficit hyperactivity disorder, nervous system disorders, and early-onset extreme obesity. In this study, whole-genome sequence was obtained from 72 founders of an intensely phenotyped experimental swine herd at the U.S. Meat Animal Research Center (USMARC) in Clay Center, Nebraska. This included 24 boars (12 Duroc and 12 Landrace ...


Detection Of Imprinted Qtl In The Berkshire X Yorkshire Cross, Jack Dekkers, Hauke Thomsen, Hakkyo Lee, Massoud Malek, Max Rothschild 2018 Iowa State University

Detection Of Imprinted Qtl In The Berkshire X Yorkshire Cross, Jack Dekkers, Hauke Thomsen, Hakkyo Lee, Massoud Malek, Max Rothschild

Max Rothschild

Genome scans have enabled the detection of regions on chromosomes that contain genes that affect economic traits, so-called quantitative trait loci (QTL). An example is the genome scan that was conducted at ISU in an F2 cross between the Berkshire and Yorkshire breeds (Malek et al. 2001a,b). This study identified many QTL related to growth performance and meat quality. But this analysis only considered QTL with a Mendelian mode of expression. This implies that an effect of the Berkshire allele on the trait was assumed to be the same whether it was inherited from the F1 sire or from ...


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