Molecular Biology Commons^™

Computational Identification Of Protein Catalytic Sites: Tests, Validation, Dan Kirshner, Jerome Nilmeier, Felice Lightstone

STAR Program Research Presentations

This project is one element of the analysis “pipeline” to characterize an organism that previously has not been well-studied. Once a protein of unknown structure has been computationally modeled (based on its sequence similarity to proteins with solved structures), then catalytic sites are identified on the model by comparison to a library of known sites. This work tested the identification algorithms with a set of proteins that have known structures and catalytic sites.

Changes In Expression Of Akt Pathway Proteins Following Treatment With Rg3 In Vitro, Kathryn Schalkoff

All Theses

To assess changes in AKT pathway signaling, a recombinant protein of the G3 domain of rat laminin-5 (rG3) that specifically binds the alpha subunit of integrins α6β1 and α6β4 expressed on cancer cells (e.g., MDA-MB-231) was produced. This recombinant protein is believed to interrupt the intracellular signaling events of the AKT pathway, causing a decrease in proliferation and survival of cells after treatment. Viability assays confirmed an apoptotic effect of rG3 on cells in a dose-dependent manner. However, data from gene expression studies of Caspase-9, GRB10, and CDKNIB proved non-conclusive that rG3 is acting upon gene expression, leading to the …

Gata-Family Transcription Factors In Magnaporthe Oryzae, Cristian F. Quispe

Department of Agronomy and Horticulture: Dissertations, Theses, and Student Research

The filamentous fungus, Magnaporthe oryzae, responsible for blast rice disease, destroys around 10-30% of the rice crop annually. Infection begins when the specialized infection structure, the appressorium, generates enormous internal turgor pressure through the accumulation of glycerol. This turgor acts on a penetration peg emerging at the base of the cell, causing it to breach the leaf surface allowing its infection.

The enzyme trehalose-6- phosphate synthase (Tps1) is a central regulator of the transition from appressorium development to infectious hyphal growth. In the first chapter we show that initiation of rice blast disease requires a regulatory mechanism involving an …

Stimulator Of Neurotropic Effects Determining The Mechanism Of Action Of The Ms-818 Compound Through Protein Identification By Affinity Chromatography And Sds-Page, Charlene Seraphina Dass

HIM 1990-2015

The MS-818 compound is used in the proliferation process of neuronal cells and many biological activities that accompany this process such as astrocyte differentiation, inhibition of neuronal apoptosis, and fraction repairs. We do know the effects of this compound, but the mechanism of action remained uncertain until now. To determine the pathway of this compound, NT2 cells were cultured and lysed to isolate the proteins. Affinity Chromatography was performed in order to immobilize the MS-818 compound to a Hi-Trap NHS column. The NT2 protein sample was injected through the column and eluted with a MS-818 concentrated, high salt content elution …

Developmental Deregulation And Tumorigenesis Inhibition In 14-3-3zeta Knockout Mouse, Jun Yang

Dissertations and Theses (Open Access)

Cancer is second leading cause of death in the United States. Improving cancer care through patient care, research, education and prevention not only saves lives, but reduces health care cost as well. Breast cancer is the most leading cause of cancer incidence and cancer related death in women of the United States. 14-3-3s are a family of conserved proteins ubiquitously expressed in all eukaryotic organisms. They form complexes with hundreds of proteins by binding to specific phospho-serine/threonine containing motifs. In this way they regulate a variety of cellular processes and are involved in many human diseases especially cancer to our …

Significance Of Increased Tissue Transglutaminase In Hormone Refractory Prostate Cancer, Amy L. Han

Dissertations and Theses (Open Access)

The progression of hormone responsive to hormone refractory prostate cancer poses a major clinical challenge in the successful treatment of prostate cancer. The hormone refractory prostate cancer cells exhibit resistance not only to castrate levels of testosterone, but also to other therapeutic modalities and hence become lethal. Currently, there is no effective treatment available for managing this cancer. These observations underscore the urgency to investigate mechanism(s) that contribute to the progression of hormone-responsive to hormone-refractory prostate cancer and to target them for improved clinical outcomes.

Tissue transglutaminase (TG2) is a multifunctional pro-inflammatory protein involved in diverse physiological processes such as …

Mechanism Of Transcriptional Suppression Of A Phytochrome A Epiallele In Arabidopsis Thaliana, Gulab D. Rangani

Graduate Theses and Dissertations

Cytosine methylation in DNA is an integral part of epigenetically controlled regulatory networks in eukaryotes. Both plants and vertebrates display DNA methylation in the gene coding region; however, its role in gene expression is not well understood. Gene promoter, on the other hand, remains largely unmethylated. Acquisition of methylation in promoter results in transcriptional suppression of the gene. The goal of this research is to study the effect of coding region methylation in gene expression using a unique gene model, phyA'. phyA' is a transcriptionally suppressed epiallele of the Arabidopsis thaliana Phytochrome A gene, which contains methylation in CG sites …

The Role Of The Androgen Receptor Cofactor P44/Wdr77 In Astrocyte Activation, Bryce H. Vincent

Dissertations and Theses (Open Access)

Astrogliosis is induced by neuronal damage and is also a pathological feature of the major aging-related neurodegenerative disorders. The mechanisms that control the cascade of astrogliosis have not been well established. In a previous study, we identified a novel androgen receptor (AR)-interacting protein (p44/WDR77) and found that it plays a critical role in the control of proliferation and differentiation of prostate epithelial cells. In the present study, we found that deletion of the p44 gene in the mouse brain caused accelerated aging with dramatic astrogliosis. The p44/WDR77 is expressed in astrocytes and loss of p44/WDR77 expression in astrocytes leads to …

The Role Of Cancer-Associated Fibroblasts In Lung Tumorigenesis, Jonathon D. Roybal

Dissertations and Theses (Open Access)

The extracellular milieu is rich in growth factors that drive tumor progression,but the mechanisms that govern tumor cell sensitivity to those ligands have notbeen fully defined. In this study, we address this question in mice that developmetastatic lung adenocarcinomas through the suppression of the microRNA-200 (miR-200) family. Cancer-associated fibroblasts (CAF) enhance tumorgrowth and invasion by secreting VEGF-A that binds to VEGFR1, a processrequired for tumor growth and metastasis in mice and correlated with a poorprognosis in lung adenocarcinoma patients. In this study, we discovered thatmiR-200 blocked CAF-induced tumor cell invasion by directly targetingVEGFR1 in tumor cells. In the context of …

Downregulation Of Pax2 Suppresses Ovarian Cancer Cell Growth, Huijuan Song

Dissertations and Theses (Open Access)

PAX2 is one of nine PAX genes regulating tissue development and cellular differentiation in embryos. PAX2 promotes cell proliferation, oncogenic transformation, cell-lineage specification, migration, and survival. Unattenuated PAX2 has been found in several cancer types. We therefore sought to elucidate the role of PAX2 in ovarian carcinomas. We found that PAX2 was expressed in low-grade serous, clear cell, endometrioid and mucinous cell ovarian carcinomas, which are relatively chemoresistant compared to high grade serous ovarian carcinomas. Four ovarian cancer cell lines, RMUGL (mucinous), TOV21G (clear cell), MDAH-2774 (endometrioid) and IGROV1 (endometrioid), which express high-levels of PAX2, were used to study the …

Dissecting The Interaction Between P53 And Trim24, Aundrietta D. Duncan

Dissertations and Theses (Open Access)

Dissecting the Interaction of p53 and TRIM24

Aundrietta DeVan Duncan

Supervisory Professor, Michelle Barton, Ph.D.

p53, the “guardian of the genome”, plays an important role in multiple biological processes including cell cycle, angiogenesis, DNA repair and apoptosis. Because it is mutated in over 50% of cancers, p53 has been widely studied in established cancer cell lines. However, little is known about the function of p53 in a normal cell. We focused on characterizing p53 in normal cells and during differentiation. Our lab recently identified a novel binding partner of p53, Tripartite Motif 24 protein (TRIM24). TRIM24 is a member of …

Mechanisms Of Adenovirus-Mediated Autophagy, Erin White

Dissertations and Theses (Open Access)

A patient diagnosed with a glioma, generally, has an average of 14 months year to live after implementation of conventional therapies such as surgery, chemotherapy, and radiation. Glioblastomas are highly lethal because of their aggressive nature and resistance to conventional therapies and apoptosis. Thus other avenues of cell death urgently need to be explored. Autophagy, which is also known as programmed cell death type II, has recently been identified as an alternative mechanism to kill apoptosis- resistant cancer cells. Traditionally, researchers have studied how cells undergo autophagy during viral infection as an immune response mechanism, but recently researchers have discovered …

Cloning, Expression, And Characterization Of The Aminoethylphosphonate Pathway In Trypanosoma Cruzi, Samuel I. Rodriguez^, Esteban M. Cordero, Igor C. Almeida*

COURI Symposium Abstracts, Summer 2011

No abstract provided.

Sunday Driver/Jip3 Binds Kinesin Heavy Chain Directly And Enhances Its Motility, Faneng Sun, Chuanmei Zhu, Ram Dixit, Valeria Cavalli

Biology Faculty Research

Neuronal development, function and repair critically depend on axonal transport of vesicles and protein complexes, which is mediated in part by the molecular motor kinesin‐1. Adaptor proteins recruit kinesin‐1 to vesicles via direct association with kinesin heavy chain (KHC), the force‐generating component, or via the accessory light chain (KLC). Binding of adaptors to the motor is believed to engage the motor for microtubule‐based transport. We report that the adaptor protein Sunday Driver (syd, also known as JIP3 or JSAP1) interacts directly with KHC, in addition to and independently of its known interaction with KLC. Using an in vitro motility assay, …

The Mitospecific Region Of Mrp20 And Its Importance For The Assembly Of Mitochondrial Ribosomes In Saccharomyces Cerevisiae, Jasvinder Kaur

Dissertations (1934 -)

Yeast mitochondrial ribosomes are composed of an rRNA scaffold, encoded by the mitochondrial genome and many different proteins, which, with the exception of one, are encoded by nuclear genes. These ribosomal proteins are imported into the mitochondrial matrix following their synthesis in the cytosol, however, little is known about the subsequent events which result in an assembled, translationally-competent ribosome. Many of the mitochondrial ribosomal proteins bear homology to bacterial ancestors. In addition to the acquisition of mitochondrial targeting signals, a number of these nuclearly-encoded ribosomal proteins have acquired additional domains, often at their C-termini, which are termed "mitochondrial-specific domains". The …

Nanosecond Pulsed Electric Field Induction Of Programmed Cell Death Is Cell Type Dependent: An In Vitro Study, Wei Ren

Theses and Dissertations in Biomedical Sciences

Nanosecond pulsed electric fields (nsPEFs) present a novel and effective method for cancer ablation by eradicating the ubiquitous cancer hallmark of apoptosis evasion and enforcing cancer programmed cell death. To develop nsPEFs as an anticancer method, a comprehensive understanding of cell death mechanisms is required. The overall objective of this dissertation is to elucidate molecular mechanisms underlying effects of nsPEFs on E4 murine squamous cell carcinoma and human T-cell Jurkat clones that are wildtype, deficient in FADD (ΔFADD) and deficient in caspase-8 (ACas-8). The overall hypothesis is that nsPEFs eliminate cancer cells through activating caspase-dependent and caspase-independent programmed cell death …

Characterization Of Rnai-Defective Mutants—Mut13-2 And Mut20—In Chlamydomonas Reinhardtii, Arit Ghosh

School of Biological Sciences: Dissertations, Theses, and Student Research

RNA interference (RNAi) is a highly conserved and sequence-specific process for regulation of gene expression. At the heart of the RNAi machinery lie 21-24 nt small non-coding RNA molecules which are able to downregulate the expression of cognate sequences, by causing degradation or translational repression of mRNAs. These noncoding RNAs act via the effector RNA-induced silencing complex (RISC), which includes core proteins such as the Argonautes (AGOs). Two Chlamydomonas reinhardtii RNAi-defective mutants, Mut13-2 and Mut20, show deletions of various domains of the Tudor Staphylococcal Nuclease 1 (TSN1) gene. TSN1 has already been recognized as a component of the RNA-induced silencing …

Identification And Characterization Of A Stilbene Synthase Gene Family In Peanut Hairy Roots, Jose Marcos Condori

Student Theses and Dissertations

Plants produce a large diversity of specialized metabolites in response to biotic and abiotic stimuli. Even though these metabolites are functional to plants as defense compounds, they also show a multitude of benefits in human health. Among these compounds are the stilbenoids. In addition to resveratrol, the most characterized of the stilbenoids, species like peanut produces several prenylated stilbenoids including arachidin-1 and arachidin-3. Recent studies suggest that these prenylated compounds have higher biological activities than resveratrol. Therefore there is a need to understand the mechanisms involved in the biosynthesis of these compounds. Some stilbene synthases involved in the biosynthesis of …

What Is New For An Old Molecule? Systematic Review And Recommendations On The Use Of Resveratrol, Ole Vang, Nihal Ahmad, Karen Brown, Anna Csiszar, Thomas Szekeres, Thomas Walle, Joseph M. Wu

NYMC Faculty Publications

BACKGROUND: Resveratrol is a natural compound suggested to have beneficial health effects. However, people are consuming resveratrol for this reason without having the adequate scientific evidence for its effects in humans. Therefore, scientific valid recommendations concerning the human intake of resveratrol based on available published scientific data are necessary. Such recommendations were formulated after the Resveratrol 2010 conference, held in September 2010 in Helsingør, Denmark.

METHODOLOGY: Literature search in databases as PUBMED and ISI Web of Science in combination with manual search was used to answer the following five questions: (1)Can resveratrol be recommended in the prevention or treatment of …

The Csc Is Required For Complete Radial Spoke Assembly And Wild-Type Ciliary Motility, Erin E. Dymek, Thomas Heuser, Daniela Nicastro, Elizabeth F. Smith

Dartmouth Scholarship

The ubiquitous calcium binding protein, calmodulin (CaM), plays a major role in regulating the motility of all eukaryotic cilia and flagella. We previously identified a CaM and Spoke associated Complex (CSC) and provided evidence that this complex mediates regulatory signals between the radial spokes and dynein arms. We have now used an artificial microRNA (amiRNA) approach to reduce expression of two CSC subunits in Chlamydomonas. For all amiRNA mutants, the entire CSC is lacking or severely reduced in flagella. Structural studies of mutant axonemes revealed that assembly of radial spoke 2 is defective. Furthermore, analysis of both flagellar beating and …