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Characterization Of The Full-Length Bag3 Protein And Stress Induced Formation Of Bag3-Z, Ahmed Gamal Abdalla Zied
Characterization Of The Full-Length Bag3 Protein And Stress Induced Formation Of Bag3-Z, Ahmed Gamal Abdalla Zied
Master's Theses
Bcl2-associated athanogene-3 (BAG3) is a pro-autophagy co-chaperone that we havepreviously shown localizes to the cardiac sarcomere and is critical for proteostasis and maintenance of normal sarcomeric function. Indeed, BAG3 loss in heart failure (HF) results in accumulation of ubiquitinated sarcomeric proteins, and depressed maximum force generating capacity (Fmax). However, how BAG3 is regulated in the cell is not well understood, with uncertainty about its structure and proteoforms. During our analysis of human heart tissue, BAG3 appears as a “doublet”, with one band at 74 kDa (BAG3-Z) and a second at a higher 85 kDa (BAG3-FL). Previous studies hypothesized the full-length …
Characterizing The Dynamic Localization Of Cmi In Early Drosophila Development, Asra Habibullah
Characterizing The Dynamic Localization Of Cmi In Early Drosophila Development, Asra Habibullah
Master's Theses
The COMPASS-like family of lysine methyltransferases, MLR/MLX complexes, are epigenetic regulators that are essential for normal development through the methylation of the fourth lysine residue on histone 3 (H3K4), a universal epigenetic mark associated with active transcription. This family of complexes is highly conserved from yeast to mammals and the genes encoding the human MLR complexes have been associated with various developmental diseases and cancers (Dingwall and Fagan, 2019). In D. melanogaster, the enzymatic methyltransferase core of this complex is composed of two proteins: Cara Mitad (Cmi, also known as Lpt) and Trithorax-related (Trr). Although these proteins have been shown …
The Impacts Of Framework Mutations On Steady-State Kinetics Of An Aldolase Abzyme, Charles Andrew Mettler
The Impacts Of Framework Mutations On Steady-State Kinetics Of An Aldolase Abzyme, Charles Andrew Mettler
Master's Theses
During affinity maturation, antibodies acquire point mutations that lead to enhanced binding strength to a particular ligand while simultaneously becoming more conformationally restricted as they become increasingly complementary to their ligand’s electrostatic geometry. This property has catalytic potential in transition-state (TS) stabilization whereby kinetic barriers (i.e., the energetic gap between the substrate and its TS) are reduced and reaction rates are enhanced. This basic mechanism is thought to underlie the majority of the rate enhancement observed for enzymes. A key integration of antibody binding specificity and strength with enzymatic catalytic strategy is the development of catalytic antibodies (“abzymes”). Abzymes are …
Function Of Mllt3 In Liver Metabolism, Grace Ann Hammack
Function Of Mllt3 In Liver Metabolism, Grace Ann Hammack
Master's Theses
The chromatin regulator MLLT3 recognizes target genes through the YEATS domain that identifies post-translationally modified histones, with preference for crotonyl and acetyl marks, and recruits different multiprotein effector complexes through its C-terminal domain to target genes. To study the role of MLLT3 in gene regulation, the Zeleznik-Le and Hemenway labs developed Mllt3 whole-body knockout (Mllt3;Rosa26-CreERT2) mice. These mice have a hematopoietic stem cell phenotype and an unexpected obesity and hepatic steatosis phenotype. It was unknown whether these phenotypes were from liver intrinsic effects or influenced by other parts of the body. To study this fatty liver phenotype further, Mllt3;Alb-Cre were …
Beta-Adrenergic Receptor Mediated Transcriptional Dysregulation In Hematopoeitc Stem And Progenitor Cells Leads To Bone Marrow Erythroid Suppresion In Multiple Myeloma Patients - Ex Vivo Investigations, Vimal Ravi Subramaniam
Beta-Adrenergic Receptor Mediated Transcriptional Dysregulation In Hematopoeitc Stem And Progenitor Cells Leads To Bone Marrow Erythroid Suppresion In Multiple Myeloma Patients - Ex Vivo Investigations, Vimal Ravi Subramaniam
Master's Theses
The sympathetic nervous system (SNS) regulates our body’s involuntary response to stress or harm. Once activated, it releases hormones and neurotransmitters known as catecholamines throughout the body to mediate its recovery. The catecholamines bind to adrenergic receptors located on various cells in a process designated as adrenergic signaling to elicit this response. This process is not always to our benefit. Adverse symptoms such as anemia can result due to adrenergic signaling. Our laboratory previously showed that burn injury triggers adrenergic receptor stimulation resulting in anemia induced by diminished erythropoiesis. The development of anemia in these studies was found to be …
Mllt3 Regulates Expression Of Genes Critical To Liver Metabolic Homeostasis, Jonathan Richard Dixit
Mllt3 Regulates Expression Of Genes Critical To Liver Metabolic Homeostasis, Jonathan Richard Dixit
Master's Theses
Hepatic steatosis (fatty liver) is increasingly prevalent in the US, and is characterized by triglyceride accumulation within hepatocytes. This is a direct result of an imbalance between lipid import, export, de novo lipogenesis, and beta oxidation. Hepatic steatosis can also progress to nonalcoholic steatohepatitis (NASH) which is characterized by increased hepatocyte damage, inflammation, and fibrosis. Further progression can lead to liver failure or hepatocellular carcinoma, requiring liver transplant or causing death. Our lab recently created Mllt3 conditional whole body knockout mice. These mice developed hepatic steatosis and NASH following Mllt3 deletion. MLLT3 is a master gene regulator that is part …
Characterizing The Requirement Of The Cmi/Trr Compass-Like Complex During Drosophila Development, Timothy Nickels
Characterizing The Requirement Of The Cmi/Trr Compass-Like Complex During Drosophila Development, Timothy Nickels
Master's Theses
The MLR family of COMPASS-like complexes are histone methyltransferase complexes that are associated with the activation of gene enhancers. In D. melanogaster, Cara mitad (Cmi, also known as Lpt) and Trithorax related (Trr) are central subunits of a complex orthologous to mammalian Lysine methyltransferase 2 C and D (KMT2C and KMT2D, also known as MLL3 and MLL2/4) that catalyze H3K4 monomethylation. Previous studies have demonstrated that mutations in these genes are associated with cancer and developmental disorders, but the mechanisms by which these alterations contribute to disease states are unknown. The Cmi-containing COMPASS-like complex and orthologous vertebrate complexes have been …
Inhibition Of Mutant Egfr In Nsclc Promotes Endothelin-1-Mediated Nsclc Disease Progression, Stephen Ollosi
Inhibition Of Mutant Egfr In Nsclc Promotes Endothelin-1-Mediated Nsclc Disease Progression, Stephen Ollosi
Master's Theses
Angiogenesis in NSCLC has been identified as important therapeutic target in combination with EGFR TKIs. However, only small incremental advancements have been made for the use of angiogenesis inhibitors in NSCLC and it remains elusive why the inhibition of VEGF-mediated neovascularization is not therapeutically efficacious. I present experimental evidence that a subpopulation of NSCLC cells with EGFR TKI-induced EMT contributes toward the attenuation of the response to EGFR TKI therapy. One of the hallmarks of cancer is heterogeneity and I have previously demonstrated that tumor heterogeneity within NSCLC cells lines harboring EGFR kinase domain mutations gives rise to divergent resistance …
Utilizing Single-Molecule Fret Methods To Study Conformational Changes In Trim5Α, Margret Suzanne Bradley
Utilizing Single-Molecule Fret Methods To Study Conformational Changes In Trim5Α, Margret Suzanne Bradley
Master's Theses
Single-molecule FRET (smFRET) is a method by which dynamic conformational changes can be monitored in a protein microscopically and in real time. smFRET relies on the creation of FRET (Förster Resonance Energy Transfer) between small molecule fluorophores conjugated to the biomolecules of interest. FRET efficiency allows calculation of interfluorophore distances. Changes in FRET efficiency represent changes in protein conformation which can inform further structural and molecular studies of the protein of interest. For example, in the Campbell Lab, we study the protein TRIM5α, an antiretroviral cellular protein which can cause premature dissociation of the HIV capsid core by an unknown …
Potential Efficacy Of Targeting Mll1 In Breast Cancer, Austin Gable Holmes
Potential Efficacy Of Targeting Mll1 In Breast Cancer, Austin Gable Holmes
Master's Theses
In 2017, it is estimated that breast cancer will be the most prevalent newly diagnosed cancer in females, at 30% in the U.S. alone1. From the early 1990s to 2014 the death rate in females from breast cancer has dropped by 10% but still stands as the second highest cancer related death in females in the U.S.2. One of the biggest hurdles in breast cancer research is disease heterogeneity. New mechanisms of disease development and progression are encountered frequently. One mechanism studied in the past decades is epigenetics. It has been theorized that the cancer epigenome can maintain an abnormal …
The Role Of C-Abl Kinase In Hcc Development, Lennox Chitsike
The Role Of C-Abl Kinase In Hcc Development, Lennox Chitsike
Master's Theses
Hepatocellular Carcinoma is the second most lethal cancer after pancreatic cancer. Unresectable HCC tumors carry a poor prognosis and few treatment options are available. The dismal prognosis is mainly due to limited therapy options and molecularly targeted therapy is deemed as solution. Here, we report a novel role of c-Abl in HCC development. We provide evidence of c-Abl activation in human HCC samples compared to normal liver. Using genetic and pharmacological tools, we show that c-Abl plays a vital role in HCC progression in vitro and in vivo. We have identified Axl as an effector in processes mediated by c-Abl. …
Role Of Pkc Delta In Uv Radiation Dna Damage Repair, Gargi Patil
Role Of Pkc Delta In Uv Radiation Dna Damage Repair, Gargi Patil
Master's Theses
DNA damage caused by ultraviolet radiation (UV), such as cyclobutane pyrimidine dimers (CPD), is repaired by the nucleotide excision repair (NER) pathway. When NER is defective, DNA damage is not repaired, leading to mutations and skin cancer. After DNA damage, the cell cycle is halted at various checkpoints to allow time for repair of the damage and maintain genomic integrity, however little is known about the coordination between NER DNA damage repair and cell cycle halting at checkpoints after DNA damage. Protein kinase C δ (PKCδ) plays major role in apoptosis and maintains the G2/M checkpoint in response to UV …
A Bioluminescence Sensor Of Nlrp3 Inflammasome Activation, Michael Alexander Winek
A Bioluminescence Sensor Of Nlrp3 Inflammasome Activation, Michael Alexander Winek
Master's Theses
The innate immune system is many organisms first line of defense against pathogenic insult or tissue damage. This defense strategy is intent on restoring homeostasis upon perturbation. Upon activation of the innate immune system in humans, an oligomeric protein complex termed the “Inflammasome” forms in myeloid cells. The canonical output of activation of any subset of inflammasome is Caspase-1-mediated secretion of pro-inflammatory cytokines IL1β and IL18. Chronic or uncontrolled inflammasome activation is at the core of myriad economically burdening diseases. In many of these diseases, endogenous factors chronically engage the innate immune system. To study these diseases in in vivo, …
Emetine As An Anti-Cancer Therapeutic In Bladder Cancer, Valerie Davidson
Emetine As An Anti-Cancer Therapeutic In Bladder Cancer, Valerie Davidson
Master's Theses
Bladder cancer is a serious health concern among the older population, as it is responsible for thousands of deaths annually in the United States. Patients that are diagnosed with muscle-invasive disease have a 5-year survival rate of only 20 percent. Additionally, muscle-invasive disease has a high metastatic potential; half of all patients develop metastatic disease within 3 years. Patients with muscle-invasive disease are presented with few treatment options aside from surgery. The current standard of care is a chemotherapeutic combination therapy of cisplatin and gemcitabine. This therapy is highly toxic, and due to the high instance of co-morbidities in these …
Determination Of An Interaction Between Nipped B-Like Protein And Mll, Adam Robert Marek
Determination Of An Interaction Between Nipped B-Like Protein And Mll, Adam Robert Marek
Master's Theses
The Mixed Lineage Leukemia (MLL) protein serves as a positive transcriptional regulator during hematopoietic and embryonic development. The MLL gene can undergo chromosomal translocations producing leukemia-causing fusions that retain the MLL amino-terminus, including the repression domain. A recent yeast two-hybrid screening used the MLL repression domain as bait and yielded nine positive clones of Nipped B-like (NIPBL).
NIPBL is a crucial member of the cohesin complex, which functions in the segregation of sister chromatids during cell division. However, recent evidence suggests the cohesin complex can also function as a transcriptional regulator.
In this study, we wanted to confirm this interaction …
Characterization Of The Coca Chemokine Receptor Four Agonist Activity Of Ubiquitin, Daniel M. Staren
Characterization Of The Coca Chemokine Receptor Four Agonist Activity Of Ubiquitin, Daniel M. Staren
Master's Theses
Ubiquitin has previously been identified as another natural agonist of CXC chemokine receptor 4 (CXCR4). In addition, recent evidence suggests that ubiquitin may activate CXCR4 through a binding site on the receptor, which is distinct from the binding site for the cognate ligand stromal cell-derived factor (SDF)-1α. The cellular consequences of ubiquitin induced CXCR4 activation, however, are still poorly defined and a side-by-side comparison of CXCR4 mediated functions after activation with SDF-1α and ubiquitin is lacking. Such information will be instrumental to better understand the physiological function of CXCR4 and to further define its role as a therapeutic target in …
Notch-1 Specifically Activates Erk1/2 In Multiple Breast Cancer Subtypes, Allison Schuyler Rogowski
Notch-1 Specifically Activates Erk1/2 In Multiple Breast Cancer Subtypes, Allison Schuyler Rogowski
Master's Theses
Notch-1 is a cell fate regulatory protein and a potent breast oncogene. Notch-1 and its ligand Jagged-1 are over-expressed in human breast cancers that are associated with poor overall survival (Reedijk, Odorcic et al. 2005). Deregulated Notch signaling may contribute to tumorigenesis by increasing proliferation, inhibiting differentiation, and preventing apoptosis (Miele, Golde et al. 2006). The mitogen-activated protein kinase (MAPK) pathway is a critical cell signaling pathway that has been implicated in the development and progression of cancer (Hanahan and Weinberg 2000). Four major MAPK pathways are involved in both cell growth and apoptosis. The regulation of these pathways is …
Wee1 Is A Biological Target Of The Mir-17-92 Cluster In Leukemia, Sonia Susan Olikara
Wee1 Is A Biological Target Of The Mir-17-92 Cluster In Leukemia, Sonia Susan Olikara
Master's Theses
MicroRNAs are noncoding RNAs that bind to the 3' untranslated region of their mRNA targets, which causes downregulation of target gene expression. Previous studies have shown that the miR-17-92 cluster, which encodes six miRNAs, is overexpressed in leukemias arising from chromosomal translocations of the Mixed Lineage Leukemia (MLL) gene. In the present study, prediction algorithms identified WEE1, a kinase that inhibits cell cycle progression, as a possible target of five of the six miRNAs. Through luciferase reporter assays, we found that miR-17, miR-20a, and miR-18a specifically target nucleotides 465 to 487 of the 3' UTR of WEE1, while miR-19a and …
Structure-Function Relationship Studies Of The Udp-Glucose Pyrophosphorylase From Escherichia Coli, Agnieszka Maria Orlof
Structure-Function Relationship Studies Of The Udp-Glucose Pyrophosphorylase From Escherichia Coli, Agnieszka Maria Orlof
Master's Theses
UDP-Glucose Pyrophosphorylase (UDP-Glc PPase) is a key enzyme of the carbohydrate metabolic pathway widely used among prokaryotes and eukaryotes. In plants, UDP-Glc PPase is necessary for sucrose synthesis while mammals utilize this enzyme for the production of glycogen. A bacterium such as Escherichia coli uses UDP-Glc PPase for biosynthesis of the lipopolysscharide core which forms a cell wall.
This study focused on UDP-Glc PPase from Escherichia coli which is encoded by both galU and galF genes. The first part of this project investigated amino acids that could play an important role in the function of UDP Glc PPase (GalU). Based …
The Distribution Of Gnrh And Erbeta In Pre- And Post-Pubertal Male Rats, Maysaa Nageeb
The Distribution Of Gnrh And Erbeta In Pre- And Post-Pubertal Male Rats, Maysaa Nageeb
Master's Theses
The hypothalamic-pituitary- gonadal axis is intrinsic in regulating reproductive function. Each part orchestrates the synthesis and release of a key hormone. At the level of the hypothalamus, Gonadotropin- releasing hormone (GnRH) is released and it is the
most upstream regulator of sexual maturation. GnRH neurons express a type of estrogen receptor, ERbeta, which suggests that estradiol (E2) plays a direct role in modulating GnRH function (Herbison and Pape 2001). The objective of this study was to determine if gonadal steroid hormones regulate the expression of ERbeta and GnRH mRNA in the male brain during pubertal development. In-situ hybridization was used …
Localization And Functional Analysis Of Plasmodium Falciparum Genes Pfl2550w And Pff0750w, Carolyn Jane Strobel
Localization And Functional Analysis Of Plasmodium Falciparum Genes Pfl2550w And Pff0750w, Carolyn Jane Strobel
Master's Theses
Malaria is a parasitic disease that causes over a million deaths worldwide each year. Understanding development through the parasite's life cycle is necessary to stop disease transmission. As the genetic basis for the crucial transition from the erythrocytic asexual cycle to gametocytogenesis is unknown, we hope to better understand this transition by studying sexual stage genes and their roles in gametocytogenesis. PFL2550w and PFF0750w are genes upregulated during gametocytogenesis that were identified by a whole-genome microarray comparing gene expression between gametocyte-producing and gametocyte-deficient strains. In this study, PFL2550w was shown to be a soluble protein that is exported from the …
The Expression Of Early Insulin Signaling Components In An Animal Model Of Insulin-Resistant Diabetes Mellitus, James A. Bonini
The Expression Of Early Insulin Signaling Components In An Animal Model Of Insulin-Resistant Diabetes Mellitus, James A. Bonini
Master's Theses
No abstract provided.
The Endocytosis Of Iodinated Protein Substrates In Cultured Human Skin Fibroblasts, Daniel P. Wilcockson
The Endocytosis Of Iodinated Protein Substrates In Cultured Human Skin Fibroblasts, Daniel P. Wilcockson
Master's Theses
No abstract provided.
A High Molecular Weight Sulfhydryl Dependent Proteinase From Bovine Spleen, Thomas J. Farrell
A High Molecular Weight Sulfhydryl Dependent Proteinase From Bovine Spleen, Thomas J. Farrell
Master's Theses
No abstract provided.
The Role Of Suppressor T Lymphocytes In Renal Allograft Rejection, Fred Barry Pearlman
The Role Of Suppressor T Lymphocytes In Renal Allograft Rejection, Fred Barry Pearlman
Master's Theses
No abstract provided.
Studies On The Mechanism Of Reaction Of Papain With N-Acetyl-L-Phenylalanyl Glycinal, A Model For An Elementary Step In Catalysis, Thresiamma Kuppy
Studies On The Mechanism Of Reaction Of Papain With N-Acetyl-L-Phenylalanyl Glycinal, A Model For An Elementary Step In Catalysis, Thresiamma Kuppy
Master's Theses
No abstract provided.
The Purification, Properties And Chemical Kinetics Of Cathepsin B1, Andrew Stanley Bajkowski
The Purification, Properties And Chemical Kinetics Of Cathepsin B1, Andrew Stanley Bajkowski
Master's Theses
No abstract provided.
Aldehydic Transition State Analogue Inhibitors Of Α-Chymotrypsin, Alexandra Constance Cheerva
Aldehydic Transition State Analogue Inhibitors Of Α-Chymotrypsin, Alexandra Constance Cheerva
Master's Theses
No abstract provided.
The Development Of An Electron Capture Gas Chromatographic Method For The Assay Of Catecholaldehydes In Tissues, Balbir Kaur Dhaliwal
The Development Of An Electron Capture Gas Chromatographic Method For The Assay Of Catecholaldehydes In Tissues, Balbir Kaur Dhaliwal
Master's Theses
No abstract provided.
The Dependency Of Brain Membrane-Bound Enzymes On Exogenous Linoleic And Linolenic Acids, Frank Joseph Spitz
The Dependency Of Brain Membrane-Bound Enzymes On Exogenous Linoleic And Linolenic Acids, Frank Joseph Spitz
Master's Theses
No abstract provided.