Virology Commons^™

Phylogenetic Investigation Of Enteric Bovine Coronavirus In Ireland Reveals Partitioning Between European And Global Strains, Lynda Gunn, P. J. Collins, M. J. O'Connell, Helen O'Shea

Department of Biological Sciences Publications

Background

Bovine coronavirus is a primary cause of neonatal calf diarrhea worldwide, and is also associated with acute diarrhea in adult cattle during the winter season. There are no reports on molecular characterization of bovine coronavirus in Ireland, and little data exists apart from serological studies.

Findings

In this study, 11 neonatal (mean age 9 days) calf BCoV strains from the south of Ireland were collected over a one year period and characterized using molecular methods. The spike gene which encodes a protein involved in viral entry, infectivity and immune response shows the most variability amongst the isolates and was …

A Genetic And Biochemical Analysis Of Sulfolobus Spindle-Shaped Virus 1, Eric Alexander Iverson

Dissertations and Theses

Viruses infecting the Archaea exhibit a tremendous amount of morphological and genetic diversity. This is especially true for crenarchaeal viruses from the family Fuselloviridae, which possess spindle-shaped capsids and genomes that harbor a great number of uncharacterized genes. The functions of these unidentified gene products are of interest as they have the potential to provide valuable insights into the fusellovirus infection cycle and archaeal viruses in general. In an effort to better characterize the genetic requirements of the Fuselloviridae, we have performed genetic and biochemical experiments using the best studied fusellovirus, Sulfolobus spindle-shaped virus 1 (SSV1).

A comprehensive …

The Evolutionary Selective Pressures Exerted On A3 Actinobacteriophages, Cheyenne Weeks-Galindo

Masters Theses, 2010-2019

This study identified evolutionary selective pressures within subcluster A3 actinobacteriophages. These phages are able to infect the clinically important genus Mycobacterium. Understanding the selective pressures on genes in these phage genomes is a step toward understanding the adaptations that result from short-term and long-term associations of phages and bacteria that have been co-evolving for perhaps billions of years. In this study 149 phamilies (phage protein families) of homologous gene sequences were analyzed using Datamonkey. Complete data were obtained for 57 phamilies. Of these, eleven phamilies were affected by recombination, three showed evidence of predominantly diversifying selection, and twenty-four have …

Expansion Of The Chlorovirus Genus By Studies On Virus Natural History And Chlorella Host Metabolism, Cristian F. Quispe

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

Inland waters cover about 2.5 percent of our planet and harbor huge numbers of known and unknown microorganisms including viruses. Viruses likely play dynamic, albeit largely undocumented roles in regulating microbial communities and in recycling nutrients in the ecosystem. Phycodnaviruses are a genetically diverse, yet morphologically similar, group of large dsDNA-containing viruses (160- to 560-kb) that inhabit aquatic environments. Members of the genus Chlorovirus are common in freshwater. They replicate in eukaryotic, single-celled, chlorella-like green algae that normally exist as endosymbionts of protists in nature. Very little is known about the natural history of the chloroviruses and how they achieve …

Structural And Functional Studies Of The Papain-Like Protease 2 From Mouse Hepatitis Virus, Yafang Chen

Open Access Dissertations

Our goal is to establish a system to investigate how the deubiquitinating (DUB) and deISGylating activities of coronavirus (CoV) papain-like protease domains (PLPs) are involved in virus immune evasion. To this end, we chose PLP2 from mouse hepatitis virus (MHV) as our target of study because MHV has historically served as a model system for the study of CoVs, and it has undeniable advantage of ease in culturing in comparison to human coronaviruses.

It is reported here the expression and purification of a region of MHV nsp3 that contains the catalytic core of the PLP2 domain and its neighboring domains. …

Understanding The Causal Agent Of Rose Rosette Disease, Patrick Louis Di Bello

Graduate Theses and Dissertations

A number viruses are known to infect roses, ranging from those in the genera Nepovirus, and Ilarvirus, which have been reported since the inception of rose virology, to recently discovered viruses in the genera Carmovirus, Closterovirus, Emaravirus, Luteovirus, Rosadnavirus, and Potyvirus. Of the viral diseases in rose, arguably the most damaging is Rose rosette (RRD), which is associated with the Emaravirus, Rose rosette virus (RRV). The objective of this thesis is to fill in the gaps in knowledge on the epidemiological aspects of RRD and RRV. There has been significant progress in the epidemiology of the RRD agent prior to …

Degradation Of The Cancer Genomic Dna Deaminase Apobec3b By Siv Vif, Allison M. Land, Jiayi Wang, Emily K. Law, Ryan Aberle, Andrea Kirmaier, Annabel Krupp, Welkin E. Johnson, Reuben S. Harris

Biological Sciences Department Publications

APOBEC3B is a newly identified source of mutation in many cancers, including breast, head/neck, lung, bladder, cervical, and ovarian. APOBEC3B is a member of the APOBEC3 family of enzymes that deaminate DNA cytosine to produce the promutagenic lesion, uracil. Several APOBEC3 family members function to restrict virus replication. For instance, APOBEC3D, APOBEC3F, APOBEC3G, and APOBEC3H combine to restrict HIV-1 in human lymphocytes. HIV-1 counteracts these APOBEC3s with the viral protein Vif, which targets the relevant APOBEC3s for proteasomal degradation. While APOBEC3B does not restrict HIV-1 and is not targeted by HIV-1 Vif in CD4-positive T cells, we asked whether related …

Detection Of Panulirus Argus Virus 1 In Caribbean Spiny Lobsters, Megan M. Montgomery-Fullerton, Roland Cooper, Kathryn M. Kauffman, Jeffrey D. Shields, Robert E. Ratzlaff

Roland A. Cooper

Panulirus argus Virus 1 (PaV1) is a pathogenic virus that infects Caribbean spiny lobsters P. argus in the Florida Keys. We have developed a PCR detection assay for PaV1 for the purpose of studying the natural history of the virus and for monitoring the prevalence of infection. The detection of the virus in hemolymph and other tissues is based on the PCR amplification of a 499 bp product using specific primers designed from a cloned fragment of the PaV1 genome. The sensitivity limit for the assay was 1.2 fg of purified viral DNA. The PaV1 primers did not react with …

Loss Of Glycosaminoglycan Receptor Binding After Mosquito Cell Passage Reduces Chikungunya Virus Infectivity, Amber M. Paul, Dhiraj Acharya, John F. Anderson, Faqing Huang, Fengwei Bai

Publications

Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that can cause fever and chronic arthritis in humans. CHIKV that is generated in mosquito or mammalian cells differs in glycosylation patterns of viral proteins, which may affect its replication and virulence. Herein, we compare replication, pathogenicity, and receptor binding of CHIKV generated in Vero cells (mammal) or C6/36 cells (mosquito) through a single passage. We demonstrate that mosquito cell derived CHIKV (CHIKVmos) has slower replication than mammalian cell derived CHIKV (CHIKVvero), when tested in both human and murine cell lines. Consistent with this, CHIKVmos infection in both cell lines produce less cytopathic …

Bacteriophages Isolated From Lake Michigan Demonstrate Broad Host-Range Across Several Bacterial Phyla, Kema Malki, Alex Kula, Katherine Bruder, Emily Sible, Thomas Hatzopoulos, Stephanie Steidel, Siobhan C. Watkins, Catherine Putonti

Biology: Faculty Publications and Other Works

BACKGROUND:

The study of bacteriophages continues to generate key information about microbial interactions in the environment. Many phenotypic characteristics of bacteriophages cannot be examined by sequencing alone, further highlighting the necessity for isolation and examination of phages from environmental samples. While much of our current knowledge base has been generated by the study of marine phages, freshwater viruses are understudied in comparison. Our group has previously conducted metagenomics-based studies samples collected from Lake Michigan - the data presented in this study relate to four phages that were extracted from the same samples.

FINDINGS:

Four phages were extracted from Lake Michigan …

A Tail Of Two Phages: Genomic And Functional Analysis Of Listeria Monocytogenes Phages Vb_Lmos_188 And Vb_Lmos_293 Reveal The Receptor-Binding Proteins Involved In Host Specificity, Aidan Casey, Kieran Jordan, Horst Neve, Aidan Coffey, Olivia Mcauliffe

Department of Biological Sciences Publications

The physical characteristics of bacteriophages establish them as viable candidates for downstream development of pathogen detection assays and biocontrol measures. To utilize phages for such purposes, a detailed knowledge of their host interaction mechanisms is a prerequisite. There is currently a wealth of knowledge available concerning Gram-negative phage-host interaction, but little by comparison for Gram-positive phages and Listeria phages in particular. In this research, the lytic spectrum of two recently isolated Listeria monocytogenes phages (vB_LmoS_188 and vB_LmoS_293) was determined, and the genomic basis for their observed serotype 4b/4e host-specificity was investigated using comparative genomics. The late tail genes of these …

Pseudomonas Bacteriophage Phi6 As A Model For Virus Emergence, Brian Elwood Ford

Dissertations, Theses, and Capstone Projects

The Pseudomonas bacteriophage Φ6 has a long and well-established history as a model organism. Here we describe a set of experiments to extend this model system to concepts previously unclaimed. Chapter 1 presents a brief background of the ecology of viruses that infect microorganisms. Chapter 2 examines genetic mutations allowing for host range expansion. Chapter 3 presents a novel paired strain assay to study how a non genetic host-acquired factor affects fitness of these enveloped viruses on subsequent hosts. Chapter 4 is an extension of this system to include how the bacteria host is affected in virus-host coevolution.

Ornithine Decarboxylase Encoded By Chlorella Virus Pbcv-1, Tiara A. Morehead,, James R. Gurnon, Byron Adams, Kenneth Nickerson, Lisa A. Fitzgerald, James L. Van Etten

Byron Adams

Sequence analysis of the 330-kb genome of chlorella virus PBCV-1 revealed an open reading frame, A207R, which encodes a protein with 37–41% amino acid identity to ornithine decarboxylase (ODC) from many eukaryotic organisms. The a207r gene was cloned and the protein was expressed as a His-A207R fusion protein in Escherichia coli. The recombinant protein catalyzes pyridoxal 5’-phosphate-dependent decarboxylation of ornithine to putrescine, the first step in the polyamine biosynthetic pathway The enzyme has a pH optimum of 9.0 and a temperature optimum of 42°C, and it requires dithiothreitol for maximal activity. The enzyme has a Km, for ornithine of 0.78 …

Characterizing The C-Terminal Region Of Human Adenovirus E1a: An Undiscovered Country, Michael J. Cohen

Electronic Thesis and Dissertation Repository

Human Adenovirus (HAdV) E1A is the first protein expressed during viral infection. The primary function of E1A is to reprogram the cell for viral replication, but it is additionally capable of transforming primary rodent cells in co-operation with other oncogenes such as HAdV E1B. Despite extensive study, little is known about the function and cellular targets of the C-terminal region of E1A. Importantly, this region is required for the transforming ability of E1A with E1B, but can also suppress transformation with Ras. Previous studies showed that interaction with the C-terminal Binding Protein (CtBP) plays a role in both functions described …

Role Of The Dna Sensor Sting In Protection From Lethal Infection Following Corneal And Intracerebral Challenge With Herpes Simplex Virus 1, Zachary M. Parker, Aisling A. Murphy, David. A. Leib

Dartmouth Scholarship

STING is a protein in the cytosolic DNA and cyclic dinucleotide sensor pathway that is critical for the initiation of innate responses to infection by various pathogens. Consistent with this, herpes simplex virus 1 (HSV-1) causes invariable and rapid lethality in STING-deficient (STING(-/-)) mice following intravenous (i.v.) infection. In this study, using real-time bioluminescence imaging and virological assays, as expected, we demonstrated that STING(-/-) mice support greater replication and spread in ocular tissues and the nervous system. In contrast, they did not succumb to challenge via the corneal route even with high titers of a virus that was routinely lethal …

Characterization Of The Replication Of Coxsackievirus B3 With A Mutationally Disrupted Cre(2c) And The Use Of Creatinine-Hydrochloride As An Antibacterial Agent, Shane E. Smithee

Theses & Dissertations

Following natural or experimental infection, and in cell culture, coxsackie B virus (CVB) RNA can persist for weeks in the absence of CPE yet with detectable viral RNA. Earlier studies in our laboratory demonstrated that this persistence produced viral RNA with up to 49 nucleotide deletions at the 5’ genomic terminus, partially degrading the cloverleaf (or domain I), an RNA structure that is required for efficient viral replication. A cis-acting replication element (CRE) in the 2C protein coding region [CRE(2C)] templates the addition of two uridine residues to the virus-encoded RNA replication primer, VPg, prior to genomic replication. Because …

Cmv Chemokines And Co-Infection: A Dissemination Plot That Peptides Can Foil, Pranay Dogra

Doctoral Dissertations

Human Cytomegalovirus (HCMV) is the leading cause of both non-hereditary mental retardation and hearing loss, and CMV infection/reactivation causes serious complications in transplant and immune compromised patients. Due to these issues, development of a CMV vaccine and/or therapeutics is required. To achieve this goal, it is necessary to gain a better understanding of CMV pathogenesis. Because of its coevolution with humans, HCMV has evolved genes with homology to human immune modulatory genes. Several of these genes help CMV establish a successful and lifelong infection within the host. An example is the viral CXC chemokine homolog UL146 gene (vCXCL-1). UL146 varies …

The Effect Of Environmental Selection Pressure On The Rate Of Recombination To An Advantageous Receptor Mutation In Bovine Coronavirus, Gavin Schroter, Sally Hall, Mona Hwang, Monica Borucki

STAR Program Research Presentations

Bovine Coronavirus (BCoV) is an important analogue in understanding the effectiveness of zoonotic, single-stranded, positive sense RNA viruses. Many of the most recent viral outbreaks have been attributed to RNA viruses that have one, or more, animal reservoirs [1]. BCoV is such a great candidate for studying these types of viruses because they are from the family Coronaviridae, which also contains the viruses that cause severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS). The goal of this study was to observe changes in genetic makeup of the virus’ outer membrane Spike protein via recombination between two …

Kaposi’S Sarcoma-Associated Herpesvirus Transmission And Infection Among Young Zambian Children, Landon Olp

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

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiologic agent for all forms of Kaposi’s sarcoma (KS)—one of the most common pediatric cancers in sub-Saharan Africa during the AIDS epidemic. KS was endemic in sub-Saharan Africa prior to the HIV/AIDS epidemic, but KS cases drastically increased thereafter. Our laboratory previously observed that KSHV infection is common among Zambian children and saliva is the major route of transmission. However, additional factors associated with the transmission of KSHV to children are poorly understood. Since a vaccine against KSHV is not currently available, it is paramount to understand factors associated with transmission so that alternative …

Preliminary Evaluation Of Near Infrared Spectroscopy As A Method To Detect Plasma Leakage In Children With Dengue Hemorrhagic Fever, Babs R. Soller, Anon Srikiatkhachorn, Fengmei Zou, Alan L. Rothman, In-Kyu Yoon, Robert V. Gibbons, Siripen Kalayanarooj, Stephen J. Thomas, Sharone Green

Sharone Green

BACKGROUND: Dengue viral infections are prevalent in the tropical and sub-tropical regions of the world, resulting in substantial morbidity and mortality. Clinical manifestations range from a self-limited fever to a potential life-threatening plasma leakage syndrome (dengue hemorrhagic fever). The objective of this study was to assess the utility of near infrared spectroscopy (NIRS) measurements of muscle oxygen saturation (SmO2) as a possible continuous measure to detect plasma leakage in children with dengue.

METHODS: Children ages 6 months to 15 years of age admitted with suspected dengue were enrolled from the dengue ward at Queen Sirikit National Institute for Child Health. …

Selective Involvement Of The Checkpoint Regulator Vista In Suppression Of B-Cell, But Not T-Cell, Responsiveness By Monocytic Myeloid-Derived Suppressor Cells From Mice Infected With An Immunodeficiency-Causing Retrovirus, Kathy A. Green, Li Wang, Randolph J. Noelle, William R. Green

Dartmouth Scholarship

Inhibition of T-cell responses in tumor microenvironments by myeloid-derived suppressor cells (MDSCs) is widely accepted. We demonstrated augmentation of monocytic MDSCs whose suppression of not only T-cell, but also B-cell, responsiveness paralleled the immunodeficiency during LP-BM5 retrovirus infection. MDSCs inhibited T cells by inducible nitric oxide synthase (iNOS)/nitric oxide (NO), but uniquely, inhibition of B cells was ~50% dependent each on iNOS/NO and the MDSC-expressed negative-checkpoint regulator VISTA. Blockade with a combination of iNOS/NO and VISTA caused additive or synergistic abrogation of MDSC-mediated suppression of B-cell responsiveness.

Functional Characterization Of P3n-Pipo Protein In The Potyviral Life Cycle, Hoda Yaghmaiean

Electronic Thesis and Dissertation Repository

Potyviruses represent the largest genus of plant-infecting viruses and include many agriculturally important viruses such as Turnip mosaic virus (TuMV), Soybean mosaic virus (SMV) and Plum pox virus (PPV). The potyviral genome consists of a large open reading frame (ORF) and a small ORF owing to a translational or transcriptional slippage in the P3 cistron. The polyproteins encoded by these two ORFs are proteolytically processed into 11 mature proteins. Recent studies have shown that P3N-PIPO, the frameshift resulting protein, is a plasmodesmata (PD)-located protein and involved in potyviral cell-to-cell movement by mediating the targeting of the potyviral CI protein to …

Chronic Inflammation As A Result Of Hepatitis C Virus Infection: A Review Of The Literature, Samantha L. Lane

DePaul Discoveries

Approximately 170 million people are infected with Hepatitis C virus (HCV) worldwide^5,6. It is estimated that roughly 80% of those infected suffer from persistent infection with the virus; this persistence of infection is progressive, and over time can lead to fibrosis, cirrhosis, and hepatocellular carcinoma⁷. Chronic inflammation and apoptotic deregulation are both hallmarks of chronic HCV infection, and many molecular pathways are initiated in both the innate and adaptive immune responses during infection with this viral pathogen. The aim of this review was to survey some of the major molecular mechanisms responsible for the induction of …

Detection Of Hcmv Viral Il-10 (Vil-10) In Healthy Blood Donors, Vivian P. Young

Master's Theses

Human cytomegalovirus (HCMV) is widespread in the general population and can establish lifelong latency with expression of a limited subset of viral genes. The UL111A gene is expressed during both lytic and latent infection, and at least two protein isoforms are produced. During lytic infection, the full length transcript yields cmvIL-10, a potent immunosuppressive viral ortholog of human IL-10 (hIL-10). Alternative splicing of the UL111A transcript yields a truncated protein, LAcmvIL-10, which is expressed during both lytic and latent infection but with a limited range of immunosuppressive functions. The two viral cytokines, collectively termed viral IL-10 (vIL-10), are identical in …

Functional Analysis Of The Molluscum Contagiosum Virus Mc160 Death Effector Domain-Containing Protein Rxdl Motif, Sarah Weber

Seton Hall University Dissertations and Theses (ETDs)

The Molluscum contagiosum virus (MCV) is a member of the Poxviridae family that causes benign skin lesions. MCV lesions persist on average for 8-12 months in otherwise healthy individuals. MCV lesions are characterized by reduced inflammation. The persistence and reduction of inflammation at the site of MCV lesions have been attributed to MCV immune evasion genes. MCV encodes two death effector domain (DED) containing proteins, MC159 and MC160. DEDs are found in cellular proteins such as FADD and procaspase-8. These cellular proteins are involved in several innate immune responses such as apoptosis and activation of interferon (IFN). Presumably, MC159 and …

Addressing The Black Box Phenomenon Of Genome Sequencing And Assembly, Brandon Carter

Senior Honors Projects, 2010-2019

Genomics, a study of all genetic material in an organism, is a new discipline having a great impact on medicine, agriculture, and environmental phenomena. Most undergraduate faculty members were not formally trained in genomics and must retool themselves in order to stay current with these evolving technologies. Advances in sequencing technology have resulted in an explosion of “big data” that can only be managed and analyzed using digital methods. Multiple complex computer programs are required to teach students the concepts using hands-on methods. These programs are challenging to use, especially since the same faculty members lacking genomics training were not …

Transduction As The Method Of Horizontal Gene Transfer Of The Staphylococcal Chromosomal Cassette Mec (Sccmec), Amber B. Sauder

Senior Honors Projects, 2010-2019

Methicillin-resistant Staphylococcus aureus (MRSA) gains resistance to β-lactam antibiotics through a mutated penicillin binding protein (PBP2a) encoded on the SCCmec element. In combination with the recombinase encoded by ccr, these two genes are used as markers of the mobile genetic element (SCCmec). Due to recent increases in community acquired MRSA infections, the mechanisms of antibiotic resistance gene transfer have gained attention. Transduction, a method of horizontal gene transfer mediated by bacteriophage, is believed to be responsible for the movement of the SCCmec element. Recent studies have shown the transduction of the SCCmec element in clinical isolates; however, this study is …

Tricistronic Lentivirus Vector Construction Using Scar-Less Dna Assembly Methods And Web-Based Software J5 To Help Study Grk4, Christophe Langouet-Astrie

Senior Honors Projects, 2010-2019

About one third of the world’s population is affected by hypertension, or high blood pressure, which increases an individual’s risk for cardiovascular disease. A major contributor to hypertension is dietary sodium intake. To assess an individual’s risk for hypertension, patients are put on a low sodium diet. However, research has shown that low salt intake can also have different and potentially harmful effects. Because of this, a genetic screen for salt sensitivity is needed to asses an individual’s salt sensitivity classification before testing and these results matched to a recommended dietary change. The kidney regulates the body’s fluid volume, so …

Effect Of Cmvil-10 On Exosome Production By Human Breast Cancer Cells, Susanna N. Basappa

Undergraduate Honors Theses

Human cytomegalovirus (HCMV) is a ubiquitous virus that infects 70-90% of the general population, primarily the immunocompromised, but has been implicated in several forms of cancer, including breast cancer. Breast cancer is the second leading cause of cancer related deaths in women in North America, usually from metastasis. Exosomes are 30-100nm vesicles produced by most cells which carry protein and RNA to cells in their microenvironment. The aim of this study is to investigate the impact of HCMV-infection of a secreted viral cytokine, cmvIL-10, on exosome production by highly metastatic breast cancer cells.

MDA-MB-231 cells were cultured in vitro, …

Elucidating The Impact Of Roseophage On Roseobacter Metabolism And Marine Nutrient Cycles, Nana Yaw Darko Ankrah

Doctoral Dissertations

As the most abundant biological entities in marine environments, viruses are an important component of marine food webs. The activity of viruses contributes significantly to the mortality of marine microorganisms, ultimately influencing biological function and chemical composition of aquatic systems by impacting species composition and flow of carbon, nitrogen and other nutrients. Despite the growing recognition that viral activity contributes to marine biogeochemical cycles, the extent to which virus infection reshapes host metabolism and the effect of this alteration on the composition of host lysate remains poorly understood. Additionally, the degree to which natural bacterioplankton communities metabolise the released lysate …