Organisms Commons^™

Crystal Structure Of A Virus-Encoded Putative Glycosyltransferase, Yi Xiang, Ulrich Baxa, Ying Zhang, Alasdair C. Steven, Gentry L. Lewis, James L. Van Etten, Michael G. Rossmann

James Van Etten Publications

The chloroviruses (family Phycodnaviridae), unlike most viruses, encode some, if not most, of the enzymes involved in the glycosylation of their structural proteins. Annotation of the gene product B736L from chlorovirus NY-2A suggests that it is a glycosyltransferase. The structure of the recombinantly expressed B736L protein was determined by X-ray crystallography to 2.3-Å resolution, and the protein was shown to have two nucleotide-binding folds like other glycosyltransferase type B enzymes. This is the second structure of a chlorovirus-encoded glycosyltransferase and the first structure of a chlorovirus type B enzyme to be determined. B736L is a retaining enzyme and belongs …

Antibiotic Sensitivity Patterns Of Hospital-Acquired And Community-Acquired Methicillin Resistant Staphylococcus Aureus, Iyad Kaddora

Theses, Dissertations and Capstones

Staphylococcus aureus is one of the most dangerous human pathogens. An intensive effort to control resistant staphylococci, especially methicillin resistant Staphylococcus aureus (MRSA), is vital as it is the most common cause of hospitalacquired infections. During the one year study period, a total of 35 MRSA isolates were collected. Fifteen isolates were identified as hospital-acquired (HA) infections, and 20 isolates were determined to be community acquired (CA). All 15 (100%) HA-MRSA strains were resistant to clindamycin and to erythromycin. Thirteen isolates (87%) were resistant to ciprofloxacin and levofloxacin, and 12 (80%) were resistant to moxifloxacin. Of the 20 CA-MRSA isolates, …

Microarray Analysis Of Paramecium Bursaria Chlorella Virus 1 Transcription, Giane M. Yanai-Balser, Garry A. Duncan, James D. Eudy, Dong Wang, Xiao Li, Irina V. Agarkova, David D. Dunigan, James L. Van Etten

James Van Etten Publications

Paramecium bursaria chlorella virus 1 (PBCV-1), a member of the family Phycodnaviridae, is a large doublestranded DNA, plaque-forming virus that infects the unicellular green alga Chlorella sp. strain NC64A. The 330-kb PBCV-1 genome is predicted to encode 365 proteins and 11 tRNAs. To monitor global transcription during PBCV-1 replication, a microarray containing 50-mer probes to the PBCV-1 365 protein-encoding genes (CDSs) was constructed. Competitive hybridization experiments were conducted by using cDNAs from poly(A)- containing RNAs obtained from cells at seven time points after virus infection. The results led to the following conclusions: (i) the PBCV-1 replication cycle is temporally …

The Isolation Of Viruses Infecting Archaea, Kenneth M. Stedman, Kate Porter, Mike L. Dyall-Smith

Biology Faculty Publications and Presentations

A mere 50 viruses of Archaea have been reported to date; these have been investigated mostly by adapting methods used to isolate bacteriophages to the unique growth conditions of their archaeal hosts. The most numerous are viruses of thermophilic Archaea. These viruses have been discovered by screening enrichment cultures and novel isolates from environmental samples for their ability to form halos of growth inhibition, or by using electron microscopy to screen enrichment cultures for virus-like particles. Direct isolation without enrichment has not yet been successful for viruses of extreme thermophiles. On the other hand, most viruses of extreme halophiles, the …

Contribution Of The Membrane-Proximal Region Of The Vesicular Stomatitis Virus Glycoprotein To Host Cell Entry And Membrane Fusion, Elizabeth Lane Matheny

Theses and Dissertations (ETD)

Vesicular stomatitis virus (VSV) is an enveloped, nonsegmented, negative-sense, RNA virus belonging to the Rhabdoviridae family. VSV is considered the prototypic Rhabdovirus due to its simple genetic organization, broad host cell tropism, and ability to be easily grown in cell culture. Therefore, VSV has been used as the prototype to study viral entry, transcription, replication, and assembly. Viral entry, a critical step in the lifecycle of the virus, is mediated by the outer surface protein, G and will be the focus of this dissertation.

We hypothesize that the highly conserved residues in the membrane-proximal region of VSV G protein are …

Manipulation Of The Moloney Murine Leukemia Virus Envelope Protein In An Effort To Develop Directly And Indirectly Targeted Retroviral Vectors For Use In Human Gene Therapy, Geneva M. Vasser

Theses and Dissertations (ETD)

Highly effective, targeted therapies against cancer would revolutionize the way people recover from this devastating illness. Gone would be the lingering side effects of the current non-specific treatments and in their place would be faster recovery times, better quality of life both during and after treatment, and less ambiguity about whether or not treatment was effective. This concept will elude modern medicine until treatments can be tailored to the patient's individual and unique disease. This concept of a transient, targeted, and tailored vehicle aimed at cancer cells lends itself to the use of replication deficient retroviral gene therapy vectors with …

Studies Of Entry, Reverse Transcription, And Regulation Of Splicing In Retroviruses, Timothy A. Sullivan

Theses and Dissertations (ETD)

The study of retroviruses and their lifecycle has contributed immensely to our knowledge of the world of biology and medicine. The central dogma of the basic flow of genetic information was shattered when the discovery that retroviruses copy their RNA genome into DNA was made. The same enzyme that performs this step, reverse transcriptase (RT), also revolutionized molecular biology when it was used as a tool to generate full length cDNA clones of expressed genes. The impact of retroviruses on the medical field has been extremely exciting as the ideas of using retroviral vectors to deliver genes providing long term …

Investigation Of The Vesicular Stomatitis Virus Matrix Protein: Uncoating And Assembly, Chad E. Mire

Theses and Dissertations (ETD)

Vesicular Stomatitis Virus (VSV) is a simple, enveloped, nonsegmented negative-strand RNA virus and is the prototype rhabdovirus to study viral entry, transcription, replication, and assembly. The matrix protein (M) of VSV is a central component of the viral replication cycle. While being the smallest of the viral proteins it is multifunctional and is involved in uncoating, cytopathic effects (CPE), and assembly of the virus. M protein interactions involved in the uncoating and assembly of VSV have been examined in this dissertation.

Uncoating of VSV involves dissociation of M from the ribonucleoprotein core (RNPs) of the virus. Current models of VSV …

Helical Peptides Derived From Lactoferrin Bind Hepatitis C Virus Envelope Protein E2, Reem Beleid, Donna Douglas, Norman Kneteman, Kamaljit Kaur

Pharmacy Faculty Articles and Research

Hepatitis C virus is a major cause of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma infecting more than 170 million people. Hepatitis C virus envelope 2 glycoprotein (E2) binds several cell‐surface molecules that act as receptor candidates mediating hepatitis C virus entry into hepatocytes. Peptides derived from human lactoferrin have been shown to bind hepatitis C virus‐E2 protein thereby preventing hepatitis C virus entry in cultured hepatocytes. In this study, starting from a 33‐residue human lactoferrin‐derived peptide, a number of biotin‐linked α‐peptides were synthesized and investigated for their E2 protein binding activity. E2 protein from hepatitis C virus genotype 1b …

Transmembrane Domain Length Of Viral K+ Channels Is A Signal For Mitochondria Targeting, Jorg Balss, Panagiotis Papatheodorou, Mario Mehmel, Dirk Baumeister, Brigitte Hertel, Nicolas Delaroque, Frank C. Chatelain, Daniel L. Minor Jr., James L. Van Etten, Joachim Rassow, Anna Moroni, Gerhard Thiel

James Van Etten Publications

K⁺ channels operate in the plasma membrane and in membranes of organelles including mitochondria. The mechanisms and topogenic information for their differential synthesis and targeting is unknown. This article describes 2 similar viral K⁺ channels that are differentially sorted; one protein (Kesv) is imported by the Tom complex into the mitochondria, the other (Kcv) to the plasma membrane. By creating chimeras we discovered that mitochondrial sorting of Kesv depends on a hierarchical combination of N- and C-terminal signals. Crucial is the length of the second transmembrane domain; extending its C terminus by >2 hydrophobic amino acids redirects Kesv …

Tetanus And Rabies Eradication In Pakistan; A Mission Not Impossible, Mohammad Wasay, Ismail A. Khatri, Naseem Salahuddin

Department of Medicine

No abstract provided.

Differential Role Of Nadp+ And Nadph In The Activity And Structure Of Gdp-D-Mannose 4,6-Dehydratase From Two Chlorella Viruses, Floriana Fruscione, Laura Sturla, Garry Duncan, James L. Van Etten, Paola Valbuzzi, Antonio De Flora, Eleonopra Di Zanni, Michela Tonetti

James Van Etten Publications

GDP-D-mannose 4,6-dehydratase (GMD) is a key enzyme involved in the synthesis of 6-deoxyhexoses in prokaryotes and eukaryotes. Paramecium bursaria chlorella virus-1 (PBCV-1) encodes a functional GMD, which is unique among characterized GMDs because it also has a strong stereospecific NADPH-dependent reductase activity leading to GDP-D-rhamnose formation (Tonetti, M., Zanardi, D., Gurnon, J., Fruscione, F., Armirotti, A., Damonte, G., Sturla, L., De Flora, A., and Van Etten, J.L. (2003) J. Biol. Chem. 278, 21559–21565). In the present study we characterized a recombinant GMD encoded by another chlorella virus, Acanthocystis turfacea chlorella virus 1 (ATCV-1), demonstrating that it has the expected dehydratase …

Downstream Pathway Activation In Cultured Hepatocytes Following Treatment With Francisella Tularensis, Nicole Wakefield

Undergraduate Theses and Capstone Projects

Francisella tularensis, the intracellular pathogen that causes tularemia, was investigated to determine how it infects and replicates within mammalian hepatocytes. To date, it has been shown that hepatocytes can be infected by F. tularensis and that this causes considerable change in the protein phosphorylation state of several vital signaling molecules within the host cell. Protein kinase pathways can be mapped as signatures of infection. Hepatocytes tend to be susceptible to infection by F. tularensis, thus stimulating internal signaling. The Francisella strains used were selected with the goal of producing a model that can be used to elucidate the cell signaling …

Structure And Function Of A Chlorella Virus Encoded Glycosyltransferase, Ying Zhang, Ye Xiang, James L. Van Etten, Michael G. Rossmann

James Van Etten Publications

Paramecium bursaria chlorella virus-1 encodes at least 5 putative glycosyltransferases that are probably involved in the synthesis of the glycan components of the viral major capsid protein. The 1.6 Å crystal structure of one of these glycosyltransferases (A64R) has a mixed α/β fold containing a central, six-stranded β-sheet flanked by α-helices. Crystal structures of A64R, complexed with UDP, CMP, or GDP, established that only UDP bound to A64R in the presence of Mn2+, consistent with its high structural similarity to glycosyltransferases which utilize UDP as the sugar carrier. The structure of the complex of A64R, UDP-glucose, and Mn2+ showed that …

Sequence And Annotation Of The 288-Kb Atcv-1 Virus That Infects An Endosymbiotic Chlorella Strain Of The Heliozoon Acanthocystis Turfacea, Lisa A. Fitzgerald, Michael V. Graves, Xiao Li, James Hartigan, Artur J.P. Pfitzner, Ella Hoffart, James L. Van Etten

James Van Etten Publications

Acanthocystis turfacea chlorella virus (ATCV-1), a prospective member of the family Phycodnaviridae, genus Chlorovirus, infects a unicellular, eukaryotic, chlorella-like green alga, Chlorella SAG 3.83, that is a symbiont in the heliozoon A. turfacea. The 288,047-bp ATCV-1 genome is the first virus to be sequenced that infects Chlorella SAG 3.83. ATCV-1 contains 329 putative protein-encoding and 11 tRNA-encoding genes. The protein-encoding genes are almost evenly distributed on both strands and intergenic space is minimal. Thirty-four percent of the viral gene products resemble entries in the public databases, including some that are unexpected for a virus. For example, these …

Chlorella Viruses Contain Genes Encoding A Complete Polyamine Biosynthetic Pathway, Sascha Baumann, Adrienne Sander, James Gurnon, Giane M. Yanai-Balser, James L. Van Etten, Markus Piotrowski

James Van Etten Publications

Two genes encoding the putative polyamine biosynthetic enzymes agmatine iminohydrolase (AIH) and N-carbamoylputrescine amidohydrolase (CPA) were cloned from the chloroviruses PBCV-1, NY-2A and MT325. They were expressed in Escherichia coli to form C-terminal (His)6-tagged proteins and the recombinant proteins were purified by Ni2+- binding affinity chromatography. The biochemical properties of the two enzymes are similar to AIH and CPA enzymes from Arabidopsis thaliana and Pseudomonas aeruginosa. Together with the previously known virus genes encoding ornithine/arginine decarboxlyase (ODC/ADC) and homospermidine synthase, the chloroviruses have genes that encode a complete set of functional enzymes that synthesize the rare polyamine homospermidine from arginine …

Chlorella Virus Mt325 Encodes Water And Potassium Channels That Interact Synergistically, Sabrina Gazzarrini, Ming Kang, Svetlana Epimashko, James L. Van Etten, Jack Dainty, Gerhard Thiel, Anna Moroni

James Van Etten Publications

Fast and selective transport of water through cell membranes is facilitated by water channels. Water channels belonging to the major intrinsic proteins (MIPs) family have been found in all three domains of life, Archaea, Bacteria, and Eukarya. Here we show that Chlorella virus MT325 has a water channel gene, aqpv1, that forms a functional aquaglyceroporin in oocytes. aqpv1 is transcribed during infection together with MT325 kcv, a gene encoding a previously undescribed type of viral potassium channel. Coexpression of AQPV1 and MT325-Kcv in Xenopus oocytes synergistically increases water transport, suggesting a possible concerted action of the two channels in …

Chlorella Viruses, Takashi Yamada, Hideki Onimatsu, James L. Van Etten

James Van Etten Publications

Chlorella viruses or chloroviruses are large, icosahedral, plaque‐forming, double‐stranded‐DNA— containing viruses that replicate in certain strains of the unicellular green alga Chlorella. DNA sequence analysis of the 330‐kbp genome of Paramecium bursaria chlorella virus 1 (PBCV‐1), the prototype of this virus family (Phycodnaviridae), predict ∼366 protein‐encoding genes and 11 tRNA genes. The predicted gene products of ∼50% of these genes resemble proteins of known function, including many that are completely unexpected for a virus. In addition, the chlorella viruses have several features and encode many gene products that distinguish them from most viruses. These products include: (1) multiple …

The Structure Of A T=169d Algal Virus, Pbcv-1, At 15Å Resolution, X. Yan, V. Bowman, N. H. Olson, James Gurnon, James L. Van Etten, Michael G. Rossmann, T. S. Baker

James Van Etten Publications

Paramecium bursaria chlorella virus 1 (PBCV-1) (genus Chlorovirus, family Phycodnaviridae), infects certain unicellular, exsymbiotic, chlorella-like green algae [1]. The PBCV-1 virion (~1x109 Da) has a linear dsDNA genome (330kbp) and contains at least 100 different proteins [1, unpublished data]. A published cryo-electron microscopy and three-dimensional (3D) image reconstruction of PBCV-1 (26Å resolution) revealed that the outer icosahedral glycoprotein capsid, with a maximum diameter of 1,900Å along the five-fold axis, consists of 1680 trimeric capsomers (trimer) and 12 pentameric capsomers (pentamer) [2]. These capsomers are arranged on a T=169d (h=7, k=8) quasi-equivalent lattice. The trimers and pentamers are organized into 20 …

Functional Implication Of The Trna Genes Encoded In The Chlorella Virus Pbcv-L Genome, Da Young Lee, Michael V. Graves, James L. Van Etten, Tae-Jin Choi

James Van Etten Publications

The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a1891192r gene showed highly biased usage of the AGA and AUA codons compared to genes encoded by E. …

Glycosyltransferases Encoded By Viruses, Nicolas Markine-Goriaynoff, Laurent Gillet, James L. Van Etten, Haralambos Korres, Naresh Verma, Alain Vanderplasschen

James Van Etten Publications

Studies of cellular biology in recent decades have highlighted the crucial roles of glycans in numerous important biological processes, raising the concept of glycomics that is now considered as important as genomics, transcriptomics and proteomics. For millions of years, viruses have been co-evolving with their hosts. Consequently, during this co-evolution process, viruses have acquired mechanisms to mimic, hijack or sabotage host processes that favour their replication, including mechanisms to modify the glycome. The importance of the glycome in the regulation of host–virus interactions has recently led to a new concept called ‘glycovirology’. One fascinating aspect of glycovirology is the study …

Can Large Dsdna-Containing Viruses Provide Information About The Minimal Genome Size Required To Support Life?, James L. Van Etten

James Van Etten Publications

The genomes of a few viruses, such as Bacillus megaterium phage G (670 kb) and the chlorella viruses (330 to 380 kb), are larger than the predicted minimal genome size required to support life (ca. 320 kb). A comparison of the 256 proteins predicted to be required for life with the putative 376 proteins encoded by chlorella virus PBCV-1, as well as those encoded by other large viruses, indicates that viruses lack many of these “essential” genes. Consequently, it is unlikely that viruses will aid in determining the minimal number and types of genes required for life. However, viruses may …

Infectious Pancreatic Necrosis (Ipn) Of Salmonid Fishes, Phillip E. Mcallister

United States Fish and Wildlife Service: Publications

Infectious pancreatic necrosis (IPN) is a viral disease principally associated with salmonids, although IPN and lPN-like viruses have been isolated from various nonsa1rnonid fish and marine invertebrates. Acute infection occurs in 1- to 4-month-old salmonids and can result in cumulative mortality approaching 100%. In contrast, fish 6 months old or older can undergo subclinical or inapparent infection, but experience no significant mortality. Survivors of the disease can become lifelong carriers of the virus. Epizootics of IPN have been reported worldwide, and several virus serotypes are recognized. The disease known as IPN was probably first described by M'Gonigle (1941) as acute …

Plant Viruses., G. D. Mclean, L. K. Price, J. A. Weir, J. M. Allen, P. Fievez, P. A. Portmann, R. J. Mclean, J. D. Sandow

Experimental Summaries - Plant Research

Clover viruses, 82ES38, 82AL47, 82MA19, 82BR19, 82BY29; 82BU5, 82HA9. Lupin virus, diseases. Barley yellow dwarf virus, 82AL46, 82AL51, 82B10, 82BA33, 82BR16, 82BR18, 82C29, 82E27, 82ES37, 82ES40, 82JE19, 82JE20, 82KA33, 82KA34, 82ABI3, 82MA18, 82MN22, 82MT34, 82NA32, 82WH28,82B8, 82MN17, 82E24, 82MT30, 82E25, 82MN18, 82MT31, 82B9, 82ABI2, 82BA31, 82C26, 82JE17, 82WH27, 82AL45, 82BR17, 82ES39, 82MA1, 82MA117, 82MT33,

Isolation And Enumeration Of Enteric Viruses From Primary Wastewater Sludge, Michael Wayne Nath

Biological Sciences Theses & Dissertations

The presence of large numbers of enteric viruses in wastewaters has resulted in concern regarding their significance and consequences for humans. Assessments of relative significance or consequences may most accurately be made following precise measurements of the range of virus types and amounts in wastewaters and sludges and after epidemiological considerations. In this study, results are presented pertaining to enteric virus isolation, concentration, and enumeration from primary wastewater sludges.

As an integral part of a comparison of isolation procedures, several anionic surfactants were examined for extraction of enteric viruses from primary wastewater sludges. Plaquing efficiencies for poliovirus type 2 and …

Oral History Interview: Josephine R. Gorby, Josephine R. Gorby

0064: Marshall University Oral History Collection

Josephine Gorby was born on February 24, 1908. She began her teaching career in a one-room school in Wimmer, McDowell County, WV. In 1931 she began teaching in Lawrence County, OH. Mrs. Gorby’s interview focuses on growing up on a farm near Proctorville, OH, her experiences as an educator, her family’s experience during World War I, and the 1937 flood of the Ohio River and its tributaries. In the audio clip provided, Mrs. Gorby discusses the impact of the 1937 flood on the school house in Athalia, OH. During her interview, she also focuses on the construction and use of …

Oral History Interview: Edward C. Jackson, Edward C. Jackson

0064: Marshall University Oral History Collection

Edward C. Jackson was born on January 17, 1911, in Canfield, Braxton County, WV. In 1926, he moved to Fenwick, Nicholas County, WV, to pursue jobs in saw mills. In his interview, Mr. Jackson describes in great detail the design and structure of the log home he lived in as a child. He focuses on growing up on a farm and places emphasis on the type of work he did and food preparation and preservation. Mr. Jackson also discusses the tight-knit community in which he was raised. In the audio clip provided, Mr. Jackson discusses working in the lumber industry. …

Comparison Of Interferon Produced In Vitro By Monkey Kidney Cells Demonstrating Interference Against Polio Ii, Coxsackie A-9, Influenza A2/Hong Kong/68, And Echo-9 Viruses, Archer Duncan Farmer Jr.

Biological Sciences Theses & Dissertations

Abstract unavailable

Experimentation To Develop Procedures To Be Used In The Investigation Of The Effects Of Restriction On The Unmodified Dna Of The Bacteriophage Lamda, John Haynes

Honors Theses

The results of experimentation approximately twenty years ago with variuos bacteriophages by Ralston and Krueger¹⁶, Anderson and Felix¹, Luria and Human¹², and by Bertani and Weigle² have shown that the host range of a given phage depends directly on the bacterial strain on which the phage has last propagated. This event came to be known as host-induced modification or host-controlled variation. As this phage host range is controlled by the host environment and can be altered from one generation to another it has, according to Luria, made microbiology "the last stronghold of Lamarckism" …