A Single Amino Acid Change Resulting In Loss Of Fluorescence Of Egfp In A Viral
Fusion Protein Confers Fitness And Growth Advantage To The Recombinant
Vesicular Stomatitis Virus,
2012
University of Nebraska-Lincoln
A Single Amino Acid Change Resulting In Loss Of Fluorescence Of Egfp In A Viral Fusion Protein Confers Fitness And Growth Advantage To The Recombinant Vesicular Stomatitis Virus, Phat X. Dinh, Debasis Panda, Phani B. Das, Subash C. Das, Anshuman Das, Asit K. Pattnaik
School of Veterinary and Biomedical Sciences: Faculty Publications
Using a recombinant vesicular stomatitis virus encoding eGFP fused in-frame with an essential viral replication protein, the phosphoprotein P, we show that during passage in culture, the virus mutates the nucleotide C289 within eGFP of the fusion protein PeGFP to A or T, resulting in R97S/C amino acid substitution and loss of fluorescence. The resultant non-fluorescent virus exhibits increased fitness and growth advantage over its fluorescent counterpart. The growth advantage of the non-fluorescent virus appears to be due to increased transcription and replication activities of the PeGFP protein carrying the R97S/C substitution. Further, our results show that the R97S/C mutation …
Amino Acid Residues In The Non-Structural Protein 1 Of Porcine Reproductive
And Respiratory Syndrome Virus Involved In Down-Regulation Of Tnf-Cx
Expression In Vitro And Attenuation In Vivo,
2012
University of Nebraska-Lincoln
Amino Acid Residues In The Non-Structural Protein 1 Of Porcine Reproductive And Respiratory Syndrome Virus Involved In Down-Regulation Of Tnf-Cx Expression In Vitro And Attenuation In Vivo, Sakthivel Subramaniam, Lalit Beura, Byungjoon Kwon, Asit K. Pattnaik, Fernando A. Osorio
School of Veterinary and Biomedical Sciences: Faculty Publications
Porcine reproductive and respiratory syndrome virus (PRRSV) suppresses tumor necrosis factor-alpha (TNF-α) production at both transcriptional and post-transcriptional levels by its non-structural proteins 1α and 1β (Nsp1α and Nsp1β). To identifY the amino acid residues responsible for this activity, we generated several alanine substitution mutants of Nsp1α and Nsp1β. Examination of the mutant proteins revealed that Nsp1α residues Gly90, Asn91 , Arg97, Argl 00 and Arg124 were necessary for TNF-α promoter suppression, whereas several amino acids spanning the entire Nsp1β ~ were found to be required for this activity. Two mutant viruses, with mutations at Nsp1α Gly90 or Nsp1β residues …
Staphylococcal Response To Oxidative Stress,
2012
University of Nebraska - Lincoln
Staphylococcal Response To Oxidative Stress, Rosmarie Gaupp, Nagender Ledala, Greg A. Somerville
School of Veterinary and Biomedical Sciences: Faculty Publications
Staphylococci are a versatile genus of bacteria that are capable of causing acute and chronic infections in diverse host species. The success of staphylococci as pathogens is due in part to their ability to mitigate endogenous and exogenous oxidative and nitrosative stress. Endogenous oxidative stress is a consequence of life in an aerobic environment; whereas, exogenous osidative and nitrosative stress are often due to the bacteria's interaction with host immune systems. To overcome the deleterious effects of oxidative and nitrosative stress, staphylococci have evolved protection, detoxification, and repair mechanisms that are controlled by a network of regulators. In this review, …
Inefficient Replication Reduces Reca-Mediated Repair Of Uvdamaged Plasmids Introduced Into Competent Escherichia Coli,
2012
Portland State University
Inefficient Replication Reduces Reca-Mediated Repair Of Uvdamaged Plasmids Introduced Into Competent Escherichia Coli, Harout Arthur Jeiranian, Charmain T. Courcelle, Justin Courcelle
Biology Faculty Publications and Presentations
Transformation of Escherichia coli with purified plasmids containing DNA damage is frequently used as a tool to characterize repair pathways that operate on chromosomes. In this study, we used an assay that allowed us to quantify plasmid survival and to compare how efficiently various repair pathways operate on plasmid DNA introduced into cells relative to their efficiency on chromosomal DNA. We observed distinct differences between the mechanisms operating on the transforming plasmid DNA and the chromosome. An average of one UV-induced lesion was sufficient to inactivate ColE1-based plasmids introduced into nucleotide excision repair mutants, suggesting an essential role for repair …
Uvrd Participation In Nucleotide Excision Repair Is Required For The Recovery Of Dna Synthesis Following Uv-Induced Damage In Escherichia Coli,
2012
Portland State University
Uvrd Participation In Nucleotide Excision Repair Is Required For The Recovery Of Dna Synthesis Following Uv-Induced Damage In Escherichia Coli, Kelley Nicole Newton, Charmain T. Courcelle, Justin Courcelle
Biology Faculty Publications and Presentations
UvrD is a DNA helicase that participates in nucleotide excision repair and several replication-associated processes, including methyl-directed mismatch repair and recombination. UvrD is capable of displacing oligonucleotides from synthetic forked DNA structures in vitro and is essential for viability in the absence of Rep, a helicase associated with processing replication forks. These observations have led others to propose that UvrD may promote fork regression and facilitate resetting of the replication fork following arrest. However, the molecular activity of UvrD at replication forks in vivo has not been directly examined. In this study, we characterized the role UvrD has in processing …
Development Of Novel Chemical Tools For Proteasome Biology & A New Approach To 1-Azaspirocyclic Ring System,
2012
University of Kentucky
Development Of Novel Chemical Tools For Proteasome Biology & A New Approach To 1-Azaspirocyclic Ring System, Lalit Kumar
Theses and Dissertations--Chemistry
The proteasome, a multiprotease complex, is clinically validated as an anticancer target by the FDA approval of bortezomib and carfilzomib for the treatment of multiple myeloma. The emergence of resistance to proteasome inhibitors however remains a major clinical challenge. Recently, distinct types of proteasomes termed ‘intermediate proteasomes’, which contain unconventional mixtures of catalytic subunits, have been implicated with drug resistance of tumor cells. In elucidating the role of intermediate proteasomes in drug resistance, a crucial step is to unequivocally determine the subunit composition of intermediate proteasomes in cells. With this in mind, the goal of the studies reported in this …
Leptin Promotes Glioblastoma ,
2012
Northern Michigan University
Leptin Promotes Glioblastoma , Johnathan Lawrence, Nicholas J. Cook, Richard A. Rovin, Robert J. Winn
Journal Articles
The hormone leptin has a variety of functions. Originally known for its role in satiety and weight loss, leptin more recently has been shown to augment tumor growth in a variety of cancers. Within gliomas, there is a correlation between tumor grade and tumor expression of leptin and its receptor. This suggests that autocrine signaling within the tumor microenvironment may promote the growth of high-grade gliomas. Leptin does this through stimulation of cellular pathways that are also advantageous for tumor growth and recurrence: antiapoptosis, proliferation, angiogenesis, and migration. Conversely, a loss of leptin expression attenuates tumor growth. In animal models …
The Effect Of Exercise On Endothelin And Endothelin Receptor Levels In Shr And Wky Male And Female Rats,
2012
Minnesota State University, Mankato
The Effect Of Exercise On Endothelin And Endothelin Receptor Levels In Shr And Wky Male And Female Rats, Lacey Stiller
All Graduate Theses, Dissertations, and Other Capstone Projects
The role of exercise as a means to modulate renal function in hypertension is unclear. However, prior studies in the laboratory of Dr. Penny Knoblich indicate that exercised female spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats excrete significantly more sodium in response to a rise in blood pressure relative to exercised male and sedentary female SHR and WKY rats. The purpose of this study was to evaluate the effects of exercise on endothelin and endothelin receptor levels in the urine and kidneys of male and female SHR and WKY rats, to elucidate potential mechanisms to explain an increase …
Cutaneous Papilloma And Squamous Cell Carcinoma Therapy Utilizing Nanosecond Pulsed Electric Fields (Nspef),
2012
Old Dominion University
Cutaneous Papilloma And Squamous Cell Carcinoma Therapy Utilizing Nanosecond Pulsed Electric Fields (Nspef), Dong Yin, Wangrong G. Yang, Jack Weissberg, Catherine B. Goff, Weikai Chen, Yoshio Kuwayama, Amanda Leiter, Hongtao Xing, Antonie Meixel, Daria Gaut, Fikret Kirkbir, David Sawcer, P. Thomas Vernier, Jonathan W. Said, Martin A. Gundersen, H. Phillip Koeffler
Bioelectrics Publications
Nanosecond pulsed electric fields (nsPEF) induce apoptotic pathways in human cancer cells. The potential therapeutic effective of nsPEF has been reported in cell lines and in xenograft animal tumor model. The present study investigated the ability of nsPEF to cause cancer cell death in vivo using carcinogen-induced animal tumor model, and the pulse duration of nsPEF was only 7 and 14 nano second (ns). An nsPEF generator as a prototype medical device was used in our studies, which is capable of delivering 7-30 nanosecond pulses at various programmable amplitudes and frequencies. Seven cutaneous squamous cell carcinoma cell lines and five …
Systematics Of Leptopelis (Anura: Arthroleptidae) From The Itombwe Plateau, Eastern Democratic Republic Of The Congo,
2012
University of Texas at El Paso
Systematics Of Leptopelis (Anura: Arthroleptidae) From The Itombwe Plateau, Eastern Democratic Republic Of The Congo, Francisco Portillo
Open Access Theses & Dissertations
Leptopelis, a genus of Central African treefrogs, includes 51 species that live in tropical forests and savannas. Currently, only two species of Leptopelis are known from the poorly explored Itombwe Plateau in eastern Democratic Republic of the Congo (DRC). Itombwe is renowned among conservationists for its rich and endemic amphibian fauna, including: Xenopus itombwensis, Chrysobatrachus cupreonitens, Laurentophryne parkeri, Hyperolius leleupi and at least three species of Arthroleptis. Evolutionary relationships of Itombwe Leptopelis were examined by sequencing two mitochondrial genes (16S: 557 bp [base pairs], cyt b: 620 bp) and one nuclear gene (RAG1: 761 bp). Results recovered strong support for …
Arachidonic Acid Signaling In Invasive And Non-Invasive Breast Cancer Cells,
2012
University of Texas at El Paso
Arachidonic Acid Signaling In Invasive And Non-Invasive Breast Cancer Cells, Debarshi Roy
Open Access Theses & Dissertations
Breast cancer is the second largest cause of cancer-related deaths in women all over the world. Epidemiological studies suggest that the consumption of high-fat diets can promote the incidence of breast cancers in both developed and developing countries. In particular, the lipid-rich diet contains arachidonic acid (AA, a C20:4 polyunsaturated fatty acid), which has been shown to be associated with tumor formation in breast tissues. Nevertheless, the actual mechanism by which AA induces the metastatic transformation and malignancy is not well understood. The goal of my dissertation, therefore, is to identify the molecules and unravel the pathways that participate in …
Hdm2 Small-Molecule Inhibitors For Therapeutic Intervention In B-Cell Lymphoma,
2012
Wayne State University
Hdm2 Small-Molecule Inhibitors For Therapeutic Intervention In B-Cell Lymphoma, Angela Sosin
Wayne State University Dissertations
Lymphomas frequently retain wild-type (wt) p53 function but overexpress HDM2, compromising p53 activity. Therefore, lymphoma is a suitable model for studying therapeutic value of disrupting HDM2-p53 association by small-molecule inhibitors (SMIs). HDM2 SMIs have been developed and are currently under various stages of preclinical and clinical investigation. This study examined various molecular mechanisms associated and biological effects of two different classes of HDM2 SMIs: the spiro-oxindoles (MI-219) and cis-imidazoline (Nutlin-3) in lymphoma cell lines and patient-derived B-lymphoma cells. Surprisingly, results revealed significant quantitative and qualitative differences between these two agents. At the molecular level, effect of Nutlin-3 was generally more …
Comparative Developmental Transcriptomics Of Echinoderms,
2012
University of South Florida
Comparative Developmental Transcriptomics Of Echinoderms, Roy Vaughn
USF Tampa Graduate Theses and Dissertations
The gastrula stage represents the point in development at which the three primary germ layers diverge. At this point the gene regulatory networks that specify the germ layers are established and the genes that define the differentiated states of the tissues have begun to be activated. These networks have been well characterized in sea urchins, but not in other echinoderms. Embryos of the brittle star Ophiocoma wendtii share a number of developmental features with sea urchin embryos, including the ingression of mesenchyme cells that give rise to an embryonic skeleton. Notable differences are that no micromeres are formed during cleavage …
Bacterial Host Interactions In Cystic Fibrosis,
2012
Institute of Technology Tallaght
Bacterial Host Interactions In Cystic Fibrosis, Máire Callaghan, Siobhan Mcclean
Articles
Chronic infection is a hallmark of cystic fibrosis (CF) and the main contributor to morbidity. Microbial infection in CF is complex, due to the number of different species that colonise the CF lung. Their colonisation is facilitated by a host response that is impaired or compromised by highly viscous mucous, zones of hypoxia and the lack of the cystic fibrosis transmembrane regulator (CFTR). Successful dominant CF pathogens combine an effective arsenal to establish infection and counter-attack the host response, together with an ability to adapt readily to an unfavourable environment. Hypermutability is common among CF pathogens facilitating adaptation and as …
Pulling Back The Veil On Stem Cell Research: The Generation Of Induced Pluripotent Stem Cells As An Alternative To Embryonic Stem Cell Research,
2012
University of Northern Iowa
Pulling Back The Veil On Stem Cell Research: The Generation Of Induced Pluripotent Stem Cells As An Alternative To Embryonic Stem Cell Research, Sara Ternes
Honors Program Theses
Stem cell research has captured the imagination of many in society, and although it holds hope for repairing the human body, curing disease, and alleviating human suffering, the effects of this pioneering technology present profound ethical and legal implications. While a general consensus exists that the use of adult stem cells is ethically and morally just, embryonic stem (ES) cell research has generated significant debate in both the scientific and legal communities. Decades of legislation and policymaking have led to the creation of a set of human subject research principles that are now well recognized and rooted in the inherent …
Lineage-Specific Differences Between Human And Simian
Immunodeficiency Virus Regulation Of Gp120 Trimer Association And
Cd4 Binding,
2012
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute
Lineage-Specific Differences Between Human And Simian Immunodeficiency Virus Regulation Of Gp120 Trimer Association And Cd4 Binding, Andrés Finzi, Beatriz Pacheco, Shi-Hua Xiang, Marie Pancera, Alon Herschhorn, Liping Wang, Xing Zeng, Anik Desormeaux, Peter D. Kwong, Joseph Sodroski
Nebraska Center for Virology: Faculty Publications
Metastable conformations of the gp120 and gp41 envelope glycoproteins of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) must be maintained in the unliganded state of the envelope glycoprotein trimer. Binding of gp120 to the primary receptor, CD4, triggers the transition to an open conformation of the trimer, promoting interaction with the CCR5 chemokine receptor and ultimately leading to gp41-mediated virus-cell membrane fusion and entry. Topological layers in the gp120 inner domain contribute to gp120-trimer association in the unliganded state and to CD4 binding. Here we describe similarities and differences between HIV-1 and SIVmac gp120. In both …
Subunit Organization Of The Membrane-Bound Hiv-1 Envelope Glycoprotein Trimer,
2012
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Department of Microbiology and Immunobiology, Harvard Medical School
Subunit Organization Of The Membrane-Bound Hiv-1 Envelope Glycoprotein Trimer, Youdong Mao, Liping Wang, Christopher Gu, Alon Herschhorn, Shi-Hua Xiang, Hillel Haim, Xinzhen Yang, Joseph Sodroski
Nebraska Center for Virology: Faculty Publications
The trimeric human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) spike is a molecular machine that mediates virus entry into host cells and is the sole target for virus-neutralizing antibodies. The mature Env spike results from cleavage of a trimeric gp160 precursor into three gp120 and three gp41 subunits. Here we describe an ~11-Å cryo-EM structure of the trimeric HIV-1 Env precursor in its unliganded state. The three gp120 and three gp41 subunits form a cage-like structure with an interior void surrounding the trimer axis. Interprotomer contacts are limited to the gp41 transmembrane region, the torus-like gp41 ectodomain, and …
Comparative Analysis Of The Fusion Efficiency
Elicited By The Envelope Glycoprotein V1–V5
Regions Derived From Human
Immunodeficiency Virus Type 1 Transmitted
Perinatally,
2012
Nankai University
Comparative Analysis Of The Fusion Efficiency Elicited By The Envelope Glycoprotein V1–V5 Regions Derived From Human Immunodeficiency Virus Type 1 Transmitted Perinatally, Hongyan Guo, Levon Abrahamyan, Chang Liu, Mackenzie Waltke, Yunqi Geng, Qimin Chen, Charles Wood, Xiaohong Kong
Nebraska Center for Virology: Faculty Publications
Understanding the properties of viruses preferentially establishing infection during perinatal transmission of human immunodeficiency virus type 1 (HIV-1) is critical for the development of effective measures to prevent transmission. A previous study demonstrated that the newly transmitted viruses (in infants) of chronically infected mother–infant pairs (MIPs) were fitter in terms of growth, which was imparted by their envelope (Env) glycoprotein V1–V5 regions, than those in the corresponding chronically infected mothers. In order to investigate whether the higher fitness of transmitted viruses was conferred by their higher entry efficiency directed by the V1–V5 regions during perinatal transmission, the fusogenicity of Env …
Glycosyl-Phosphatidylinositol (Gpi)-Anchored Membrane Association Of The Porcine
Reproductive And Respiratory Syndrome Virus Gp4 Glycoprotein And Its Co-Localization
With Cd163 In Lipid Rafts,
2012
University of Illinois at Urbana-Champaign
Glycosyl-Phosphatidylinositol (Gpi)-Anchored Membrane Association Of The Porcine Reproductive And Respiratory Syndrome Virus Gp4 Glycoprotein And Its Co-Localization With Cd163 In Lipid Rafts, Yijun Du, Asit K. Pattnaik, Cheng Song, Dongwan Yoo, Gang Li
School of Veterinary and Biomedical Sciences: Faculty Publications
The porcine reproductive and respiratory syndrome virus (PRRSV) glycoprotein 4 (GP4) resembles a typical type I membrane protein in its structure but lacks a hydrophilic tail at the C-terminus, suggesting that GP4 may be a lipid-anchored membrane protein. Using the human decay-accelerating factor (DAF; CD55), a known glycosyl-phosphatidylinositol (GPI) lipid-anchored protein, chimeric constructs were made to substitute the GPI-anchor domain of DAF with the putative lipid-anchor domain of GP4, and their membrane association and lipase cleavage were determined in cells. The DAF-GP4 fusion protein was transported to the plasma membrane and was cleaved by phosphatidylinositol-specific phospholipase C (PI-PLC), indicating that …
Fusion Of A Fluorescent Protein To The Pul25
Minor Capsid Protein Of Pseudorabies Virus
Allows Live-Cell Capsid Imaging With Negligible
Impact On Infection,
2012
Northwestern University Feinberg School of Medicine
Fusion Of A Fluorescent Protein To The Pul25 Minor Capsid Protein Of Pseudorabies Virus Allows Live-Cell Capsid Imaging With Negligible Impact On Infection, Kevin P. Bohannon, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith
School of Veterinary and Biomedical Sciences: Faculty Publications
In order to resolve the location and activity of submicroscopic viruses in living cells, viral proteins are often fused to fluorescent proteins (FPs) and visualized by microscopy. In this study, we describe the fusion of FPs to three proteins of pseudorabies virus (PRV) that allowed imaging of capsids in living cells. Included in this study are the first recombinant PRV strains expressing FP–pUL25 fusions based on a design applied to herpes simplex virus type 1 by Homa and colleagues. The properties of each reporter virus were compared in both in vitro and in vivo infection models. PRV strains expressing FP–pUL25 …
