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Cell Biology

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Electronic Thesis and Dissertation Repository

2016

Articles 1 - 14 of 14

Full-Text Articles in Life Sciences

The Association Between Oxidative Stress, Cellular Differentiation And Galectins In Human Promyelocytic Leukemia Cells (Hl-60), James R. Vinnai Dec 2016

The Association Between Oxidative Stress, Cellular Differentiation And Galectins In Human Promyelocytic Leukemia Cells (Hl-60), James R. Vinnai

Electronic Thesis and Dissertation Repository

Galectins are a group of β-galactoside-binding proteins involved in different cellular processes including stress responses and differentiation. The role and expression of galectins under oxidative stress and during neutrophilic differentiation was examined in HL-60 cells. Galectin gene (LGALS), and galectin protein expression were determined using RT-qPCR and immunoblotting, respectively. Neutrophilic differentiation was measured via a spectrofluorometric assay. DNA methylation and JNK signaling were investigated as galectin regulatory mechanisms. Menadione-induced oxidative stress, DMSO-induced differentiation, DNA hypomethylation and JNK signaling all promoted similar galectin expression profiles. Antioxidant N-acetylcysteine attenuated the menadione-induced galectin expression but only partially attenuated DMSO-induced galectin expression. …

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Evidence Of A Role For G Protein Signaling Modulator 3 In Cell Division, Drew C. Wallace Dec 2016

Evidence Of A Role For G Protein Signaling Modulator 3 In Cell Division, Drew C. Wallace

Electronic Thesis and Dissertation Repository

Components of G protein-mediated signaling are associated with positioning and orienting the mitotic spindle in the process of cell division. However, a functional role for G protein signaling modulator 3 (GPSM3) in cell division has yet to be defined. The purpose of this study was to investigate a potential role for GPSM3 in cell division. Vascular smooth muscle cells (VSMCs) from Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats, that are known to express GPSM3, were used as a model system. Here I report that GPSM3 mRNA and protein levels varied during different stages of the cell cycle in SHR VSMCs. …

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Insights Into The Role Of Connexin26 And Connexin30 In Skin Health, Syndromic Disease, And Cutaneous Wound Healing, Eric R. Press Dec 2016

Insights Into The Role Of Connexin26 And Connexin30 In Skin Health, Syndromic Disease, And Cutaneous Wound Healing, Eric R. Press

Electronic Thesis and Dissertation Repository

Connexin26 (Cx26) and Cx30 facilitate gap junctional intercellular communication (GJIC) in the epidermis and are linked to several syndromic skin diseases. We investigated 5 disease- linked Cx26 mutants and demonstrated that the severity and extent of disease can be predicted from the gain- or loss-of function properties of each mutant, as well as the ability to induce cell death. We also used transgenic mice expressing S17F Cx26 (linked to keratitis-ichthyosis- deafness syndrome) or A88V Cx30 (linked to Clouston Syndrome) to investigate the pathophysiology these skin diseases. We demonstrated that S17F Cx26, but not A88V Cx30, promotes palmoplantar keratoderma by disrupting …

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Characterization Of The Catalytic Ck2 Subunits With Substitutions At Residues Involved In Inhibitor Binding, Paul Desormeaux Sep 2016

Characterization Of The Catalytic Ck2 Subunits With Substitutions At Residues Involved In Inhibitor Binding, Paul Desormeaux

Electronic Thesis and Dissertation Repository

CK2 is a constitutively active, ubiquitously expressed and pleiotropic serine/threonine protein kinase that is implicated in many cellular functions including tumorigenesis. CK2 has two catalytic subunits, CK2a and CK2a’, that carry out its function in the cell. Previous studies have indicated that inhibitor-refractory mutants have been effective in recovering residual CK2 activity, in the presence of inhibitors, when compared to wild type CK2. Based on these observations, inhibitor-refractory mutants were created for both CK2a and CK2a’ and tested with various concentrations with two CK2-specific inhibitors, CX-4945 and inhibitor VIII. The CK2a triple mutant (V66A/I174A/H160D) was tested in inducible U2OS Flp-In …

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Cal And Magi Pdz Protein Regulation Of Crfr1 And 5-Ht2ar Trafficking And Signaling, Maha Mahmoud Hammad Aug 2016

Cal And Magi Pdz Protein Regulation Of Crfr1 And 5-Ht2ar Trafficking And Signaling, Maha Mahmoud Hammad

Electronic Thesis and Dissertation Repository

PDZ (PSD95/Disc Large/Zona Occludens) domain-containing proteins are scaffolding proteins that play important roles in regulating the activity of G protein-coupled receptors. Corticotropin Releasing Factor Receptor 1 (CRFR1) and Serotonin 2A Receptor (5-HT2AR) are two GPCRs that are commonly associated with mental disorders. Both receptors also contain a class I PDZ-binding motif at the carboxyl terminal tail. In the first chapter, we investigate the effects of CAL (CFTR-associated ligand) on regulating the trafficking and signaling of CRFR1. We demonstrate a role for CAL in inhibiting CRFR1 endocytosis, cell surface expression, and CRF-mediated ERK1/2 signaling via the CRFR1 PDZ-binding motif. …

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The Differential Effects Of Connexin26 And Connexin43 In Modulating Epidermal Health In Response To Uv Injury, Katanya C. Alaga Aug 2016

The Differential Effects Of Connexin26 And Connexin43 In Modulating Epidermal Health In Response To Uv Injury, Katanya C. Alaga

Electronic Thesis and Dissertation Repository

Using two disease-linked connexin mutant mice that possess significantly reduced connexin function (Cx43I130T/+ and Cx26K14-S17F/+) this investigation is the first to address the roles of Cx26 and Cx43 in modulating epidermal health in response to UV radiation. Viable Cx26K14-S17F/+ mice were successfully created in-house to express the mutant Cx26 S17F protein in keratinocytes and possess scaly skin, rapidly develop epidermal desquamation, and die soon after UV exposure. Conversely, Cx43I130T/+ mice did not possess any skin abnormalities before or after UV exposure. We also identified that primary murine melanocytes are not homocellularly coupled and do not …

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Panx1 And Panx3 Regulate Adipocyte Development And Fat Accumulation In Vivo, Vanessa R. Lee Aug 2016

Panx1 And Panx3 Regulate Adipocyte Development And Fat Accumulation In Vivo, Vanessa R. Lee

Electronic Thesis and Dissertation Repository

The obesity epidemic is a growing concern due to its various comorbidities and associated risk factors. Pannexins 1 and 3 (Panx1 and Panx3), are members of a family of channel-forming glycoproteins that have been reported to be important in paracrine signaling and development. Panx1 and Panx3 are homologous and are regulated in many different cell types, mediating cell proliferation and differentiation. We have shown that Panx1 and Panx3 are expressed in adipocytes and adipose-derived stromal cells (ASCs) throughout the process of differentiation. Mice globally lacking Panx1 (Panx1 KO) have significantly greater total fat mass compared to wildtype (WT) mice under …

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Nucleoporin-Mediated Regulation Of The Kcnq1ot1 Imprinted Domain, Saqib Sachani Aug 2016

Nucleoporin-Mediated Regulation Of The Kcnq1ot1 Imprinted Domain, Saqib Sachani

Electronic Thesis and Dissertation Repository

Genomic imprinting is an epigenetic phenomenon that restricts gene expression to one parental allele while the other copy is silent. How this duality is regulated is not fully understood. Using the Kcnq1ot1 imprinted domain as a model, previous work in the laboratory identified nucleoporin 107 as a candidate regulator of imprinted domain regulation. Within the Kcnq1ot1 domain resides the imprinting control region, the paternally expressed Kcnq1ot1 (Kcnq1 opposite transcript 1) noncoding RNA, nine maternal-expressed protein-coding genes, as well as genes that escape imprint regulation. On the maternal allele, the Kcnq1ot1 imprinting control region is methylated, silencing the embedded Kcnq1ot1 …

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Inhibition Of Mt1-Mmp Proteolytic Function And Erk1/2 Signalling Influences Breast Cancer Cell Migration And Invasion Through Changes In Mmp-2 And Mmp-9 Expression, Caitlin L. Evered Aug 2016

Inhibition Of Mt1-Mmp Proteolytic Function And Erk1/2 Signalling Influences Breast Cancer Cell Migration And Invasion Through Changes In Mmp-2 And Mmp-9 Expression, Caitlin L. Evered

Electronic Thesis and Dissertation Repository

Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a multifunctional protease that invokes changes extracellularly via cleavages of ECM substrates, and intracellularly through induction of cell signalling cascades, both to influence cell behaviour and motility. The effects of MT1-MMP activation, MMP catalytic activity, and ERK1/2 signalling were examined by chemically or genetically altering each parameter. Regardless of treatment, expression of the two gelatinases, MMP-2 and -9, were always altered in an inverse relationship. This work proposes a pathway through active MT1-MMP initiation of ERK1/2 phosphorylation and subsequent targeting the NF-κB transcription factor, to ultimately influence MMP-9 expression. Modulation of MT1-MMP activation, …

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Cardiovascular Characterization Of Cx40/Panx1 Single And Double Knockout Mice, Meghan L. Jelen Jul 2016

Cardiovascular Characterization Of Cx40/Panx1 Single And Double Knockout Mice, Meghan L. Jelen

Electronic Thesis and Dissertation Repository

Connexins (Cxs) and pannexins (Panxs) are protein families that form large-pore channels which exist at the plasma membrane for both intracellular and extracellular signaling. Given their potential for overlapping cellular signaling functions we proposed that mice lacking both a connexin and a pannexin would have a severe phenotype. To investigate this possibility we crossed Panx1 null mice with Cx40 knockout mice and characterized the first global connexin/pannexin double knockout mouse. Intriguingly, the combined ablation of both Cx40 and Panx1 caused decreased prenatal and newborn survival, but did not affect the fertility or lifespan of surviving mice. Cx40-/- and Cx40 …

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Characterization Of Numb As A Regulator Of Anaplastic Lymphoma Kinase, Ran Wei Jun 2016

Characterization Of Numb As A Regulator Of Anaplastic Lymphoma Kinase, Ran Wei

Electronic Thesis and Dissertation Repository

Cellular events rely on protein-protein interactions that are often mediated by modular domains which recognize particular sequence motifs in binding partners. The NUMB protein is the first described cell fate determinant and multifaceted adaptor that is involved in a wide variety of cellular events. NUMB mainly mediates protein interactions via its modular PTB domain. Here we present a systematic investigation of the NUMB-PTB interactome by employing an integrative strategy combining both protein and peptide arrays. We profiled NUMB-PTB binding specificity and interacting proteins genome-wide. The receptor tyrosine kinases (RTKs) are found highly enriched in the interactome, raising the possibility that …

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The Role Of Nadph Oxidase In Ros Mediated Differentiation, Benjamin J. Dickson Jun 2016

The Role Of Nadph Oxidase In Ros Mediated Differentiation, Benjamin J. Dickson

Electronic Thesis and Dissertation Repository

Mouse teratocarcinoma F9 cells differentiate into primitive endoderm (PrE) when treated with retinoic acid (RA) or H2O2 and these changes are accompanied by an upregulation of Wnt6 and activation of the canonical WNT/β-catenin pathway. Data from our lab shows PrE differentiation is accompanied by an increase in reactive oxygen species (ROS), which induces a conformational change in Nucleoredoxin preventing its ability to bind and inhibit Dishevelled. This in turn positively impacts on the WNT/β- catenin signaling pathway leading to differentiation. The source of endogenous ROS seen following RA treatment was investigated and members of the NADPH oxidase (NOX) family were …

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Oncolytic Virus Therapy For The Treatment Of Metastatic Ovarian Cancer, Jessica Tong Apr 2016

Oncolytic Virus Therapy For The Treatment Of Metastatic Ovarian Cancer, Jessica Tong

Electronic Thesis and Dissertation Repository

The management of patients with epithelial ovarian cancer (EOC) faces two major challenges which standard treatments fail to effectively address: 1) Diffuse metastasis as a consequence of late stage diagnosis and 2) intra-tumoral heterogeneity, which fuels tumor evolution and drives the acquisition of chemotherapeutic resistance. In this thesis, we tested new therapeutic strategies using a 3-dimensional in vitro spheroid culture model that mimics key steps of epithelial ovarian cancer metastasis; and another model that mimics both temporal and cellular heterogeneity by establishing multiple cell lines from a single patient over the course of disease progression. Using these models, we investigated …

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A Genetic Test Of A Model For Two Activities Of Fushi Tarazu Protein In Drosophila Melanogaster, Alaa Briek Apr 2016

A Genetic Test Of A Model For Two Activities Of Fushi Tarazu Protein In Drosophila Melanogaster, Alaa Briek

Electronic Thesis and Dissertation Repository

The important pair-rule segmentation gene fushi tarazu (ftz) encodes a homeodomain (HD)-containing protein involved in the establishment of even-numbered parasegments during embryonic development. The D. melanogaster ftz is a derived homeotic selector (Hox) gene which lost its homeotic function during the evolution of arthropods. Genetic analyses have shown that FTZ has two distinct activities required during development: HD-dependent and HD-independent FTZ activities. The aim of this study was to test the interaction of the two FTZ activities proposed by Hyduk and Percival-Smith (1996), by generating site-specific mutant ftz alleles for intragenic complementation. CRISPR-mediated homology directed repair (HDR) was used to …

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