Plant Pathology Commons^™

Mono- And Digalactosyldiacylglycerol Lipids Function Nonredundantly To Regulate Systemic Acquired Resistance In Plants, Qing-Ming Gao, Keshun Yu, Ye Xia, M. B. Shine, Caixia Wang, Duroy Navarre, Aardra Kachroo, Pradeep Kachroo

Plant Pathology Faculty Publications

The plant galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) have been linked to the anti-inflammatory and cancer benefits of a green leafy vegetable diet in humans due to their ability to regulate the levels of free radicals like nitric oxide (NO). Here, we show that DGDG contributes to plant NO as well as salicylic acid biosynthesis and is required for the induction of systemic acquired resistance (SAR). In contrast, MGDG regulates the biosynthesis of the SAR signals azelaic acid (AzA) and glycerol-3-phosphate (G3P) that function downstream of NO. Interestingly, DGDG is also required for AzA-induced SAR, but MGDG is not. Notably, …

Co-Opted Oxysterol-Binding Orp And Vap Proteins Channel Sterols To Rna Virus Replication Sites Via Membrane Contact Sites, Daniel Barajas, Kai Xu, Isabel Fernández De Castro Martín, Zsuzsanna Sasvari, Federica Brandizzi, Cristina Risco, Peter D. Nagy

Plant Pathology Faculty Publications

Viruses recruit cellular membranes and subvert cellular proteins involved in lipid biosynthesis to build viral replicase complexes and replication organelles. Among the lipids, sterols are important components of membranes, affecting the shape and curvature of membranes. In this paper, the tombusvirus replication protein is shown to co-opt cellular Oxysterol-binding protein related proteins (ORPs), whose deletion in yeast model host leads to decreased tombusvirus replication. In addition, tombusviruses also subvert Scs2p VAP protein to facilitate the formation of membrane contact sites (MCSs), where membranes are juxtaposed, likely channeling lipids to the replication sites. In all, these events result in redistribution and …

A Novel Partitivirus That Confers Hypovirulence On Plant Pathogenic Fungi, Xueqiong Xiao, Jiasen Cheng, Jinghua Tang, Yanping Fu, Daohong Jiang, Timothy S. Baker, Said A. Ghabrial, Jiatao Xie

Plant Pathology Faculty Publications

Members of the family Partitiviridae have bisegmented double-stranded RNA (dsRNA) genomes and are not generally known to cause obvious symptoms in their natural hosts. An unusual partitivirus, Sclerotinia sclerotiorum partitivirus 1 (SsPV1/WF-1), conferred hypovirulence on its natural plant-pathogenic fungal host, Sclerotinia sclerotiorum strain WF-1. Cellular organelles, including mitochondria, were severely damaged. Hypovirulence and associated traits of strain WF-1 and SsPV1/WF-1 were readily cotransmitted horizontally via hyphal contact to different vegetative compatibility groups of S. sclerotiorum and interspecifically to Sclerotinia nivalis and Sclerotinia minor. S. sclerotiorum strain 1980 transfected with purified SsPV1/WF-1 virions also exhibited hypovirulence and associated traits similar …

Tombusvirus-Yeast Interactions Identify Conserved Cell-Intrinsic Viral Restriction Factors, Zsuzsanna Sasvari, Paulina Alatriste Gonzalez, Peter D. Nagy

Plant Pathology Faculty Publications

To combat viral infections, plants possess innate and adaptive immune pathways, such as RNA silencing, R gene and recessive gene-mediated resistance mechanisms. However, it is likely that additional cell-intrinsic restriction factors (CIRF) are also involved in limiting plant virus replication. This review discusses novel CIRFs with antiviral functions, many of them RNA-binding proteins or affecting the RNA binding activities of viral replication proteins. The CIRFs against tombusviruses have been identified in yeast (Saccharomyces cerevisiae), which is developed as an advanced model organism. Grouping of the identified CIRFs based on their known cellular functions and subcellular localization in yeast …

The Hop-Like Stress-Induced Protein 1 Cochaperone Is A Novel Cell-Intrinsic Restriction Factor For Mitochondrial Tombusvirus Replication, Kai Xu, Jing-Yi Lin, Peter D. Nagy

Plant Pathology Faculty Publications

Recent genome-wide screens reveal that the host cells express an arsenal of proteins that inhibit replication of plus-stranded RNA viruses by functioning as cell-intrinsic restriction factors of viral infections. One group of cell-intrinsic restriction factors against tombusviruses contains tetratricopeptide repeat (TPR) domains that directly interact with the viral replication proteins. In this paper, we find that the TPR domain-containing Hop-like stress-inducible protein 1 (Sti1p) cochaperone selectively inhibits the mitochondrial membrane-based replication of Carnation Italian ringspot tombusvirus (CIRV). In contrast, Sti1/Hop does not inhibit the peroxisome membrane-based replication of the closely related Tomato bushy stunt virus (TBSV) or Cucumber necrosis virus …

Cell Type-Dependent Rna Recombination Frequency In The Japanese Encephalitis Virus, Wei-Wei Chiang, Chingkai Chuang, Mei Chao, Wei-June Chen

Plant Pathology Faculty Publications

Japanese encephalitis virus (JEV) is one of approximately 70 flaviviruses, frequently causing symptoms involving the central nervous system. Mutations of its genomic RNA frequently occur during viral replication, which is believed to be a force contributing to viral evolution. Nevertheless, accumulating evidences show that some JEV strains may have actually arisen from RNA recombination between genetically different populations of the virus. We have demonstrated that RNA recombination in JEV occurs unequally in different cell types. In the present study, viral RNA fragments transfected into as well as viral RNAs synthesized in mosquito cells were shown not to be stable, especially …

Noncanonical Role For The Host Vps4 Aaa+ Atpase Escrt Protein In The Formation Of Tomato Bushy Stunt Virus Replicase, Daniel Barajas, Isabel Fernández De Castro Martín, Judit Pogany, Cristina Risco, Peter D. Nagy

Plant Pathology Faculty Publications

Assembling of the membrane-bound viral replicase complexes (VRCs) consisting of viral- and host-encoded proteins is a key step during the replication of positive-stranded RNA viruses in the infected cells. Previous genome-wide screens with Tomato bushy stunt tombusvirus (TBSV) in a yeast model host have revealed the involvement of eleven cellular ESCRT (endosomal sorting complexes required for transport) proteins in viral replication. The ESCRT proteins are involved in endosomal sorting of cellular membrane proteins by forming multiprotein complexes, deforming membranes away from the cytosol and, ultimately, pinching off vesicles into the lumen of the endosomes. In this paper, we show an …

The Expanding Functions Of Cellular Helicases: The Tombusvirus Rna Replication Enhancer Co-Opts The Plant Eif4aiii-Like Atrh2 And The Ddx5-Like Atrh5 Dead-Box Rna Helicases To Promote Viral Asymmetric Rna Replication, Nikolay Kovalev, Peter D. Nagy

Plant Pathology Faculty Publications

Replication of plus-strand RNA viruses depends on recruited host factors that aid several critical steps during replication. Several of the co-opted host factors bind to the viral RNA, which plays multiple roles, including mRNA function, as an assembly platform for the viral replicase (VRC), template for RNA synthesis, and encapsidation during infection. It is likely that remodeling of the viral RNAs and RNA-protein complexes during the switch from one step to another requires RNA helicases. In this paper, we have discovered a second group of cellular RNA helicases, including the eIF4AIII-like yeast Fal1p and the DDX5-like Dbp3p and the orthologous …

Characterization Of Glomerella Strains Recovered From Anthracnose Lesions On Common Bean Plants In Brazil, Quélen L. Barcelos, Joyce M.A. Pinto, Lisa J. Vaillancourt, Elaine A. Souza

Plant Pathology Faculty Publications

Anthracnose caused by Colletotrichum lindemuthianum is an important disease of common bean, resulting in major economic losses worldwide. Genetic diversity of the C. lindemuthianum population contributes to its ability to adapt rapidly to new sources of host resistance. The origin of this diversity is unknown, but sexual recombination, via the Glomerella teleomorph, is one possibility. This study tested the hypothesis that Glomerella strains that are frequently recovered from bean anthracnose lesions represent the teleomorph of C. lindemuthianum. A large collection of Glomerella isolates could be separated into two groups based on phylogenetic analysis, morphology, and pathogenicity to beans. Both …

Inactivation Of The Host Lipin Gene Accelerates Rna Virus Replication Through Viral Exploitation Of The Expanded Endoplasmic Reticulum Membrane, Chingkai Chuang, Daniel Barajas, Jun Qin, Peter D. Nagy

Plant Pathology Faculty Publications

RNA viruses take advantage of cellular resources, such as membranes and lipids, to assemble viral replicase complexes (VRCs) that drive viral replication. The host lipins (phosphatidate phosphatases) are particularly interesting because these proteins play key roles in cellular decisions about membrane biogenesis versus lipid storage. Therefore, we examined the relationship between host lipins and tombusviruses, based on yeast model host. We show that deletion of PAH1 (phosphatidic acid phosphohydrolase), which is the single yeast homolog of the lipin gene family of phosphatidate phosphatases, whose inactivation is responsible for proliferation and expansion of the endoplasmic reticulum (ER) membrane, facilitates robust RNA …

Construction Of A Sonchus Yellow Net Virus Minireplicon: A Step Toward Reverse Genetic Analysis Of Plant Negative-Strand Rna Viruses, Uma Ganesan, Jennifer N. Bragg, Min Deng, Sharon Marr, Mi Yeon Lee, Shasha Qian, Manling Shi, Justin Kappel, Cole Peters, Yeon Lee, Michael M. Goodin, Ralf G. Dietzgen, Zhenghe Li, Andrew O. Jackson

Plant Pathology Faculty Publications

Reverse genetic analyses of negative-strand RNA (NSR) viruses have provided enormous advances in our understanding of animal viruses over the past 20 years, but technical difficulties have hampered application to plant NSR viruses. To develop a reverse genetic approach for analysis of plant NSR viruses, we have engineered Sonchus yellow net nucleorhabdovirus (SYNV) minireplicon (MR) reporter cassettes for Agrobacterium tumefaciens expression in Nicotiana benthamiana leaves. Fluorescent reporter genes substituted for the SYNV N and P protein open reading frames (ORFs) exhibited intense single-cell foci throughout regions of infiltrated leaves expressing the SYNV MR derivatives and the SYNV nucleocapsid (N), phosphoprotein …

Tall Fescue Endophyte Effects On Tolerance To Water-Deficit Stress, Padmaja Nagabhyru, Randy D. Dinkins, Constance L. Wood, Charles W. Bacon, Christopher L. Schardl

Plant Pathology Faculty Publications

BACKGROUND: The endophytic fungus, Neotyphodium coenophialum, can enhance drought tolerance of its host grass, tall fescue. To investigate endophyte effects on plant responses to acute water deficit stress, we did comprehensive profiling of plant metabolite levels in both shoot and root tissues of genetically identical clone pairs of tall fescue with endophyte (E+) and without endophyte (E-) in response to direct water deficit stress. The E- clones were generated by treating E+ plants with fungicide and selectively propagating single tillers. In time course studies on the E+ and E- clones, water was withheld from 0 to 5 days, during which …

Currencies Of Mutualisms: Sources Of Alkaloid Genes In Vertically Transmitted Epichloae, Christopher L. Schardl, Carolyn A. Young, Juan Pan, Simona Florea, Johanna E. Takach, Daniel G. Panaccione, Mark L. Farman, Jennifer S. Webb, Jolanta Jaromczyk, Nikki D. Charlton, Padmaja Nagabhyru, Li Chen, Chong Shi, Adrian Leuchtmann

Plant Pathology Faculty Publications

The epichloae (Epichloë and Neotyphodium species), a monophyletic group of fungi in the family Clavicipitaceae, are systemic symbionts of cool-season grasses (Poaceae subfamily Poöideae). Most epichloae are vertically transmitted in seeds (endophytes), and most produce alkaloids that attack nervous systems of potential herbivores. These protective metabolites include ergot alkaloids and indole-diterpenes (tremorgens), which are active in vertebrate systems, and lolines and peramine, which are more specific against invertebrates. Several Epichloë species have been described which are sexual and capable of horizontal transmission, and most are vertically transmissible also. Asexual epichloae are mainly or exclusively vertically transmitted, and many are …

Three-Dimensional Structure Of Victorivirus Hvv190s Suggests Coat Proteins In Most Totiviruses Share A Conserved Core, Sarah E. Dunn, Hua Li, Giovanni Cardone, Max L. Nibert, Said A. Ghabrial, Timothy S. Baker

Plant Pathology Faculty Publications

Double-stranded (ds)RNA fungal viruses are currently assigned to six different families. Those from the family Totiviridae are characterized by nonsegmented genomes and single-layer capsids, 300-450 Å in diameter. Helminthosporium victoriae virus 190S (HvV190S), prototype of recently recognized genus Victorivirus, infects the filamentous fungus Helminthosporium victoriae (telomorph: Cochliobolus victoriae), which is the causal agent of Victoria blight of oats. The HvV190S genome is 5179 bp long and encompasses two large, slightly overlapping open reading frames that encode the coat protein (CP, 772 aa) and the RNA-dependent RNA polymerase (RdRp, 835 aa). To our present knowledge, victoriviruses uniquely express their RdRps via …

Plant-Symbiotic Fungi As Chemical Engineers: Multi-Genome Analysis Of The Clavicipitaceae Reveals Dynamics Of Alkaloid Loci, Christopher L. Schardl, Carolyn A. Young, Uljana Hesse, Stefan G. Amyotte, Kalina Andreeva, Patrick J. Calie, Damien J. Fleetwood, David C. Haws, Neil Moore, Birgitt Oeser, Daniel G. Panaccione, Kathryn K. Schweri, Christine R. Voisey, Mark L. Farman, Jerzy W. Jaromczyk, Bruce A. Roe, Donal M. O'Sullivan, Barry Scott, Paul Tudzynski, Zhiqiang An, Elissaveta G. Arnaoudova, Charles T. Bullock, Nikki D. Charlton, Li Chen, Murray Cox, Randy D. Dinkins, Simona Florea, Anthony E. Glenn, Anna Gordon, Ulrich Güldener, Daniel R. Harris, Walter Hollin, Jolanta Jaromczyk, Richard D. Johnson, Anar K. Khan, Eckhard Leistner, Adrian Leuchtmann, Chunjie Li, Jinge Liu, Jinze Liu, Miao Liu, Wade Mace, Caroline Machado, Padmaja Nagabhyru, Juan Pan, Jan Schmid, Koya Sugawara, Ulrike Steiner, Johanna E. Takach, Eiji Tanaka, Jennifer S. Webb, Ella V. Wilson, Jennifer L. Wiseman, Ruriko Yoshida, Zheng Zeng

Plant Pathology Faculty Publications

The fungal family Clavicipitaceae includes plant symbionts and parasites that produce several psychoactive and bioprotective alkaloids. The family includes grass symbionts in the epichloae clade (Epichloë and Neotyphodium species), which are extraordinarily diverse both in their host interactions and in their alkaloid profiles. Epichloae produce alkaloids of four distinct classes, all of which deter insects, and some-including the infamous ergot alkaloids-have potent effects on mammals. The exceptional chemotypic diversity of the epichloae may relate to their broad range of host interactions, whereby some are pathogenic and contagious, others are mutualistic and vertically transmitted (seed-borne), and still others vary in pathogenic …

Mites In The Promoters Of Effector Genes Allow Prediction Of Novel Virulence Genes In Fusarium Oxysporum, Sarah M. Schmidt, Petra M. Houterman, Ines Schreiver, Lisong Ma, Stefan G. Amyotte, Biju Chellappan, Sjef Boeren, Frank L. W. Takken, Martijn Rep

Plant Pathology Faculty Publications

BACKGROUND: The plant-pathogenic fungus Fusarium oxysporum f.sp.lycopersici (Fol) has accessory, lineage-specific (LS) chromosomes that can be transferred horizontally between strains. A single LS chromosome in the Fol4287 reference strain harbors all known Fol effector genes. Transfer of this pathogenicity chromosome confers virulence to a previously non-pathogenic recipient strain. We hypothesize that expression and evolution of effector genes is influenced by their genomic context.

RESULTS: To gain a better understanding of the genomic context of the effector genes, we manually curated the annotated genes on the pathogenicity chromosome and identified and classified transposable elements. Both retro- and DNA transposons are present …

Discovery Of Novel Dsrna Viral Sequences By In Silico Cloning And Implications For Viral Diversity, Host Range And Evolution, Huiquan Liu, Yanping Fu, Jiatao Xie, Jiasen Cheng, Said A. Ghabrial, Guoqing Li, Xianhong Yi, Daohong Jiang

Plant Pathology Faculty Publications

Genome sequence of viruses can contribute greatly to the study of viral evolution, diversity and the interaction between viruses and hosts. Traditional molecular cloning methods for obtaining RNA viral genomes are time-consuming and often difficult because many viruses occur in extremely low titers. DsRNA viruses in the families, Partitiviridae, Totiviridae, Endornaviridae, Chrysoviridae, and other related unclassified dsRNA viruses are generally associated with symptomless or persistent infections of their hosts. These characteristics indicate that samples or materials derived from eukaryotic organisms used to construct cDNA libraries and EST sequencing might carry these viruses, which were not easily detected by the researchers. …

Evolutionary Genomics Of Mycovirus-Related Dsrna Viruses Reveals Cross-Family Horizontal Gene Transfer And Evolution Of Diverse Viral Lineages, Huiquan Liu, Yanping Fu, Jiatao Xie, Jiasen Cheng, Said A. Ghabrial, Guoqing Li, Youliang Peng, Xianhong Yi, Daohong Jiang

Plant Pathology Faculty Publications

BACKGROUND: Double-stranded (ds) RNA fungal viruses are typically isometric single-shelled particles that are classified into three families, Totiviridae, Partitiviridae and Chrysoviridae, the members of which possess monopartite, bipartite and quadripartite genomes, respectively. Recent findings revealed that mycovirus-related dsRNA viruses are more diverse than previously recognized. Although an increasing number of viral complete genomic sequences have become available, the evolution of these diverse dsRNA viruses remains to be clarified. This is particularly so since there is little evidence for horizontal gene transfer (HGT) among dsRNA viruses.

RESULTS: In this study, we report the molecular properties of two novel dsRNA mycoviruses that …

A Co-Opted Dead-Box Rna Helicase Enhances Tombusvirus Plus-Strand Synthesis, Nikolay Kovalev, Judit Pogany, Peter D. Nagy

Plant Pathology Faculty Publications

Replication of plus-strand RNA viruses depends on recruited host factors that aid several critical steps during replication. In this paper, we show that an essential translation factor, Ded1p DEAD-box RNA helicase of yeast, directly affects replication of Tomato bushy stunt virus (TBSV). To separate the role of Ded1p in viral protein translation from its putative replication function, we utilized a cell-free TBSV replication assay and recombinant Ded1p. The in vitro data show that Ded1p plays a role in enhancing plus-strand synthesis by the viral replicase. We also find that Ded1p is a component of the tombusvirus replicase complex and Ded1p …

The Tpr Domain In The Host Cyp40-Like Cyclophilin Binds To The Viral Replication Protein And Inhibits The Assembly Of The Tombusviral Replicase, Jing-Yi Lin, Venugopal Mendu, Judit Pogany, Jun Qin, Peter D. Nagy

Plant Pathology Faculty Publications

Replication of plus-stranded RNA viruses is greatly affected by numerous host-coded proteins acting either as susceptibility or resistance factors. Previous genome-wide screens and global proteomics approaches with Tomato bushy stunt tombusvirus (TBSV) in a yeast model host revealed the involvement of cyclophilins, which are a large family of host prolyl isomerases, in TBSV replication. In this paper, we identified those members of the large cyclophilin family that interacted with the viral replication proteins and inhibited TBSV replication. Further characterization of the most effective cyclophilin, the Cyp40-like Cpr7p, revealed that it strongly inhibits many steps during TBSV replication in a cell-free …

Synergistic Roles Of Eukaryotic Translation Elongation Factors 1bγ And 1a In Stimulation Of Tombusvirus Minus-Strand Synthesis, Zsuzsanna Sasvari, Lara Izotova, Terri Goss Kinzy, Peter D. Nagy

Plant Pathology Faculty Publications

Host factors are recruited into viral replicase complexes to aid replication of plus-strand RNA viruses. In this paper, we show that deletion of eukaryotic translation elongation factor 1Bgamma (eEF1Bγ) reduces Tomato bushy stunt virus (TBSV) replication in yeast host. Also, knock down of eEF1Bγ level in plant host decreases TBSV accumulation. eEF1Bγ binds to the viral RNA and is one of the resident host proteins in the tombusvirus replicase complex. Additional in vitro assays with whole cell extracts prepared from yeast strains lacking eEF1Bγ demonstrated its role in minus-strand synthesis by opening of the structured 3' end of the viral …

Sag101 Forms A Ternary Complex With Eds1 And Pad4 And Is Required For Resistance Signaling Against Turnip Crinkle Virus, Shifeng Zhu, Rae-Dong Jeong, Srivathsa C. Venugopal, Ludmila Lapchyk, Duroy Navarre, Aardra Kachroo, Pradeep Kachroo

Plant Pathology Faculty Publications

EDS1, PAD4, and SAG101 are common regulators of plant immunity against many pathogens. EDS1 interacts with both PAD4 and SAG101 but direct interaction between PAD4 and SAG101 has not been detected, leading to the suggestion that the EDS1-PAD4 and EDS1-SAG101 complexes are distinct. We show that EDS1, PAD4, and SAG101 are present in a single complex in planta. While this complex is preferentially nuclear localized, it can be redirected to the cytoplasm in the presence of an extranuclear form of EDS1. PAD4 and SAG101 can in turn, regulate the subcellular localization of EDS1. We also show that the Arabidopsis genome …

Widespread Horizontal Gene Transfer From Circular Single-Stranded Dna Viruses To Eukaryotic Genomes, Huiquan Liu, Yanping Fu, Bo Li, Xiao Yu, Jiatao Xie, Jiasen Cheng, Said A. Ghabrial, Guoqing Li, Xianhong Yi, Daohong Jiang

Plant Pathology Faculty Publications

BACKGROUND: In addition to vertical transmission, organisms can also acquire genes from other distantly related species or from their extra-chromosomal elements (plasmids and viruses) via horizontal gene transfer (HGT). It has been suggested that phages represent substantial forces in prokaryotic evolution. In eukaryotes, retroviruses, which can integrate into host genome as an obligate step in their replication strategy, comprise approximately 8% of the human genome. Unlike retroviruses, few members of other virus families are known to transfer genes to host genomes.

RESULTS: Here we performed a systematic search for sequences related to circular single-stranded DNA (ssDNA) viruses in publicly available …

An Effective Virus-Based Gene Silencing Method For Functional Genomics Studies In Common Bean, Claudia Díaz-Camino, Padmanaban Annamalai, Federico Sanchez, Aardra Kachroo, Said A. Ghabrial

Plant Pathology Faculty Publications

BACKGROUND: Common bean (Phaseolus vulgaris L.) is a crop of economic and nutritious importance in many parts of the world. The lack of genomic resources have impeded the advancement of common bean genomics and thereby crop improvement. Although concerted efforts from the "Phaseomics" consortium have resulted in the development of several genomic resources, functional studies have continued to lag due to the recalcitrance of this crop for genetic transformation.

RESULTS: Here we describe the use of a bean pod mottle virus (BPMV)-based vector for silencing of endogenous genes in common bean as well as for protein expression. This BPMV-based vector …

Abundant Degenerate Miniature Inverted-Repeat Transposable Elements In Genomes Of Epichloid Fungal Endophytes Of Grasses, Damien J Fleetwood, Anar K Khan, Richard D Johnson, Carolyn A Young, Shipra Mittal, Ruth E Wrenn, Uljana Hesse, Simon J Foster, Christopher L. Schardl, Barry Scott

Plant Pathology Faculty Publications

Miniature inverted-repeat transposable elements (MITEs) are abundant repeat elements in plant and animal genomes; however, there are few analyses of these elements in fungal genomes. Analysis of the draft genome sequence of the fungal endophyte Epichloë festucae revealed 13 MITE families that make up almost 1% of the E. festucae genome, and relics of putative autonomous parent elements were identified for three families. Sequence and DNA hybridization analyses suggest that at least some of the MITEs identified in the study were active early in the evolution of Epichloë but are not found in closely related genera. Analysis of MITE integration …

Making Of Viral Replication Organelles By Remodeling Interior Membranes, Zsuzsanna Sasvari, Peter D. Nagy

Plant Pathology Faculty Publications

Positive-stranded RNA (+RNA) viruses exploit host cell machinery by subverting host proteins and membranes and altering cellular pathways during infection. To achieve robust replication, some +RNA viruses, such as poliovirus (PV), build special intracellular compartments, called viral replication organelles. A recent work from the Altan-Bonnett laboratory [1] gave new insights into the formation of poliovirus replication organelles, which are unique subcellular structures containing many individual replication complexes as a result of dynamic cellular membrane remodeling.

Translation Elongation Factor 1a Facilitates The Assembly Of The Tombusvirus Replicase And Stimulates Minus-Strand Synthesis, Zhenghe Li, Judit Pogany, Steven Tupman, Anthony M. Esposito, Terri Goss Kinzy, Peter D. Nagy

Plant Pathology Faculty Publications

Replication of plus-strand RNA viruses depends on host factors that are recruited into viral replicase complexes. Previous studies showed that eukaryotic translation elongation factor (eEF1A) is one of the resident host proteins in the highly purified tombusvirus replicase complex. Using a random library of eEF1A mutants, we identified one mutant that decreased and three mutants that increased Tomato bushy stunt virus (TBSV) replication in a yeast model host. Additional in vitro assays with whole cell extracts prepared from yeast strains expressing the eEF1A mutants demonstrated several functions for eEF1A in TBSV replication: facilitating the recruitment of the viral RNA template …

The Combined Effect Of Environmental And Host Factors On The Emergence Of Viral Rna Recombinants, Hannah M. Jaag, Peter D. Nagy

Plant Pathology Faculty Publications

Viruses are masters of evolution due to high frequency mutations and genetic recombination. In spite of the significance of viral RNA recombination that promotes the emergence of drug-resistant virus strains, the role of host and environmental factors in RNA recombination is poorly understood. Here we report that the host Met22p/Hal2p bisphosphate-3'-nucleotidase regulates the frequency of viral RNA recombination and the efficiency of viral replication. Based on Tomato bushy stunt virus (TBSV) and yeast as a model host, we demonstrate that deletion of MET22 in yeast or knockdown of AHL, SAL1 and FRY1 nucleotidases/phosphatases in plants leads to increased TBSV recombination …

Distinct Evolutionary Histories Of The Dna-A And Dna-B Components Of Bipartite Begomoviruses, Rob W. Briddon, Basavaprabhu L. Patil, Basavaraj Bagewadi, Muhammad Shah Nawaz-Ul-Rehman, Claude M. Fauquet

Plant Pathology Faculty Publications

BACKGROUND: Viruses of the genus Begomovirus (family Geminiviridae) have genomes consisting of either one or two genomic components. The component of bipartite begomoviruses known as DNA-A is homologous to the genomes of all geminiviruses and encodes proteins required for replication, control of gene expression, overcoming host defenses, encapsidation and insect transmission. The second component, referred to as DNA-B, encodes two proteins with functions in intra- and intercellular movement in host plants. The origin of the DNA-B component remains unclear. The study described here was initiated to investigate the relationship between the DNA-A and DNA-B components of bipartite begomoviruses with a …

Systematic Overrepresentation Of Dna Termini And Underrepresentation Of Subterminal Regions Among Sequencing Templates Prepared From Hydrodynamically Sheared Linear Dna Molecules, Sherri L. Schwartz, Mark L. Farman

Plant Pathology Faculty Publications

BACKGROUND: Analysis of fungal genome sequence assemblies reveals that telomeres are poorly represented even though telomeric reads tend to be superabundant. We surmised that the problem might lie in the DNA shearing conditions used to create clone libraries for genome sequencing.

RESULTS: A shotgun strategy was used to sequence and assemble circular and linear cosmid DNAs sheared using conditions typical for a genome project. The DNA sheared in circular form assembled into a single sequence contig. However, the linearized cosmid produced an incomplete assembly because the two DNA termini, though greatly overrepresented in the clone library used for sequencing, were …