Plant Pathology Commons^™

A Virus-Encoded Potassium Ion Channel Is A Structural Protein In The Chlorovirus Paramecium Bursaria Chlorella Virus 1 Virion, Giulia Romani, Adrianna Piotrowski, Stefan Hillmer, James Gurnon, James L. Van Etten, Anna Morani, Gerhard Thiel, Brigitte Hertel

James Van Etten Publications

Most chloroviruses encode small K+ channels, which are functional in electrophysiological assays. The experimental finding that initial steps in viral infection exhibit the same sensitivity to channel inhibitors as the viral K+ channels has led to the hypothesis that the channels are structural proteins located in the internal membrane of the virus particles. This hypothesis was questioned recently because proteomic studies failed to detect the channel protein in virions of the prototype chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1). Here, we used a mAb raised against the functional K+ channel from chlorovirus MA-1D to search for the viral K+ channel …

Potassium Ion Channels: Could They Have Evolved From Viruses?1[W], Gerhard Thiel, Anna Moroni, Guillaume Blanc, James L. Van Etten

James Van Etten Publications

Cells communicate among themselves by electrical activity. Sophisticated membrane-embedded proteins, called ion channels, catalyze rapid, selective, and regulated ion fluxes across membranes (Hille, 2001). The resulting membrane currents are responsible for neuronal activity and the systemic propagation of electrical signals in animals. The activity of some channels is important for muscle movement in animals or growth in plants; other channels sense the concentration of physiological signals and modulate key processes in all kinds of eukaryotic cells. Among the many diverse ion channels in higher organisms, K+ channels are among the most important. One feature of K+ channels is that they …

Minimal Art: Or Why Small Viral K+ Channels Are Good Tools For Understanding Basic Structure And Function Relations, Gerhard Thiel, Dirk Baumeister, Indra Schroeder, Stefan M. Kast, James L. Van Etten, Anna Moroni

James Van Etten Publications

Some algal viruses contain genes that encode proteins with the hallmarks of K+ channels. One feature of these proteins is that they are less than 100 amino acids in size, which make them truly minimal for a K+ channel protein. That is, they consist of only the pore module present in more complex K+ channels. The combination of miniature size and the functional robustness of the viral K+ channels make them ideal model systems for studying how K+ channels work. Here we summarize recent structure/function correlates from these channels, which provide insight into functional properties such as gating, pharmacology and …

Functional Hak/Kup/Kt-Like Potassium Transporter Encoded By Chlorella Viruses, Timo Greiner, José Ramos, Maria C. Alvarez, James Gurnon, Ming Kang, James L. Van Etten, Anna Moroni, Gerhard Thiel

James Van Etten Publications

Chlorella viruses are a source of interesting membrane transport proteins. Here we examine a putative K+ transporter encoded by virus FR483 and related chlorella viruses. The protein shares sequence and structural features with HAK/KUP/KT-like K+ transporters from plants, bacteria and fungi. Yeast complementation assays and Rb+ uptake experiments show that the viral protein, termed HAKCV (high-affinity K+ transporter of chlorella virus), is functional, with transport characteristics that are similar to those of known K+ transporters. Expression studies revealed that the protein is expressed as an early gene during viral replication, and proteomics data indicate that it is not packaged in …

Giant Viruses, James L. Van Etten

James Van Etten Publications

The common view of viruses, mostly true, is of tiny burglars that sneak into cells, grab the biosynthetic controls and compel the cell to make huge numbers of progeny that break out of the cell and keep the replication cycle going. Viruses are supposed to be diminutive even compared to cells that are just a micrometer (1,000 nanometers) in diameter. They are supposed to travel light, making do with just a few well-adapted genes.

In 1992, a new microorganism was isolated from a power-plant cooling tower in Bradford, England, where Timothy Robotham, a microbiologist at Leeds Public Health Laboratory, was …

A Functional Calcium-Transporting Atpase Encoded By Chlorella Viruses, Maria Cristina Bonza, Holger Martin, Ming Kang, Gentry L. Lewis, Timo Greiner, Sonia Giacometti, James L. Van Etten, Maria Ida De Michelis, Gerhard Thiel, Anna Moroni

James Van Etten Publications

Calcium-transporting ATPases (Ca²⁺ pumps) are major players in maintaining calcium homeostasis in the cell and have been detected in all cellular organisms. Here, we report the identification of two putative Ca²⁺ pumps, M535L and C785L, encoded by chlorella viruses MT325 and AR158, respectively, and the functional characterization of M535L. Phylogenetic and sequence analyses place the viral proteins in group IIB of P-type ATPases even though they lack a typical feature of this class, a calmodulin-binding domain. A Ca²⁺ pump gene is present in 45 of 47 viruses tested and is transcribed during virus infection. Complementation analysis of …

Identification Of An L-Rhamnose Synthetic Pathway In Two Nucleocytoplasmic Large Dna Viruses, Madhu Parakkottil Chothi, Garry A. Duncan, Andrea Armirotti, Chantal Abergel, James R. Gurnon, James L. Van Etten, Cinzia Bernardi, Gianluca Damonte, Michela Tonetti

James Van Etten Publications

Nucleocytoplasmic large DNA viruses (NCLDVs) are characterized by large genomes that often encode proteins not commonly found in viruses. Two species in this group are Acanthocystis turfacea chlorella virus 1 (ATCV-1) (family Phycodnaviridae, genus Chlorovirus) and Acanthamoeba polyphaga mimivirus (family Mimiviridae), commonly known as mimivirus. ATCV-1 and other chlorovirus members encode enzymes involved in the synthesis and glycosylation of their structural proteins. In this study, we identified and characterized three enzymes responsible for the synthesis of the sugar L-rhamnose: two UDP-D-glucose 4,6-dehydratases (UGDs) encoded by ATCV-1 and mimivirus and a bifunctional UDP-4-keto-6-deoxy-D-glucose epimerase/reductase (UGER) from mimivirus. Phylogenetic …

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 …

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 …

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 …

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 …

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 …

Characterization Of A PorosTm-Fumonisin B1 Affinity Column For Isolating Ceramide Synthase From Rat Liver, S. Jernigan, W. B. Melchior Jr., G. R. Jenkins, K. L. Rowland, D. W. Roberts, P. C. Howard, H. Tolleson

Journal of the Arkansas Academy of Science

Fumonisin B1 is a mycotoxin produced by fungi of the genus Fusarium, common pathogens of corn and other grain plants. Toxic effects associated with fumonisin B1 include equine leukoencephalomacia, porcine pulmonary edema, rat renal carcinoma, and murine hepatocellular carcinoma. Increased risk for esophageal cancer in humans has been epidemiologically associated with consumption of corn contaminated with Fusarium, suggesting that fumonisin B1 may be involved. The biological effects of fumonisin B1 exposure result primarily from disruption of de novo sphingolipid biosynthesis via inhibition of ceramide synthase. Exposure of animals or cultured cells to fumonisin B1 results in the characteristic accumulation of …

Bulletin 4406a - Fungal Diseases Of Canola In Western Australia, Martin Barbetti, Ravjit Khangura

Bulletins 4000 -

GRDC - Grains Research & Development Corporation.

There has been dramatic growth in the canola industry in Western Australia, from 35,000 hectares in 1993 to 920,000 hectares in 1999. Along with this expansion in the area sown the incidence and severity of various fungal diseases have also increased, especially blackleg. This Bulletin describes the most important and most prevalent fungal diseases of canola in Western Australia.

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 …

Role Of Endophytes In Tall Fescue, E. L. Piper, C. P. West

Journal of the Arkansas Academy of Science

Tall fescue (Festuca arundinacea Schreb.) is the most commonly grown cool season grass used for pastures in Arkansas. Most tall fescue contains a fungal endophyte (Acremonium coenophialum Morgan-Jones & Gams), which causes fescue toxicosis in livestock and costs cattle producers millions of dollars annually in lost production. Endophyte presence is known to reduce wild mammal populations in areas where tall fescue is prevalent. The endophyte spends its entire life cycle within the plant and is transmitted through the seed. The association is mutualistic with the plant providing nutrients for the endophyte and the endophyte conferring drought, insect, and nematode resistance …

Evaluation Of Aposphaeria Amaranthi As A Bioherbicide For Pigweed (Amaranthus Spp.), A. S. Mintz, G. J. Weidmann

Journal of the Arkansas Academy of Science

Studies were conducted to determine the potential of the fungus, Aposphaeria amaranth!, as a bioherbicide for pigweeds (Amaranthus spp.). Experiments to establish the environmental parameters necessary for control of tumble pigweed (A. albus) demonstrated that an 8-hr dew period was sufficient for control of seedlings with four to six leaves, and that temperatures ranging from 20 to 28 C were conducive for disease development. Conidial concentrations as lowas 1x 10s conidia per ml also were sufficient for plant mortality. Host range tests demonstrated pathogenicity of A. amaranthi to several other species of Amaranthus, including biotypes resistant to triazine herbicides. Disease …

Bulletin 4133 - Fungal Diseases Of Pasture Legumes In Western Australia, M. J. Barbetti

Bulletins 4000 -

Subterranean clover (Trifolium subterraneum L.) is the most important annual pasture legume in Western Australia, forming the basis of some 6.5 million ha of annual pastures. The success of subterranean clover may be attributed to its ability to increase soil fertility as a consequence of nitrogen fixed, to withstand heavy grazing, to control soil erosion and, with some cultivars, to tolerate waterlogging. Fungal diseases have been a major limitation to the productivity of subterranean clover, affecting both the amount of plant matter and the quantity of seed produced.

Annual medic (Medicago spp., in particular M. polymorpha L.) is also an …

Bulletin No 3863 - Plants Which Cause Kimberley Horse Disease, A. L. Payne

Bulletins - 3000 - 3999

Crotalaria crispata and Crotalaria retusa (Rattlepod), two plants responsible for Kimberley horse disease*, are, unfortunately, fairly widespread throughout the Kimberleys. Both species are apparently toxic in all stages of growth.

This bulletin is to assist pastoralists to:-

• Identify the two plants;

• Recognise the type of country and conditions under which they grow; and

• Locate horse paddocks on country where the plants are least likely to occur.

A Study Of Puccinia Graminis And Cronartium Ribicola, Duane W. Hughes

Graduate Student Research Papers

It was the purpose of this study (1) to review the literary history of rusts in the United States and in the Northwest in particular, ( 2) to learn where rusts generally are found geographically, (3) to gain further information regarding the etiology of wheat rust (Puccinia graminis tritici), of white pine blister rust (Cronartium ribicola) and to become enlightened on other rusts, (4) to learn ways of controlling the two rusts, black stem rust of wheat and white pine blister rust, and to gain an appreciation of the economic importance of rust prevention.

List Of The Smut Fungi Of Arkansas, George E. Templeton

Journal of the Arkansas Academy of Science

No abstract provided.