Biochemistry, Biophysics, and Structural Biology Commons^™

Pseudorabies Virus Fast Axonal Transport Occurs By A Pus9- Independent Mechanism, Gina R. Daniel, Patricia J. Sollars, Gary E. Pickard, Gregory A. Smith

School of Veterinary and Biomedical Sciences: Faculty Publications

Reactivation from latency results in transmission of neurotropic herpesviruses from the nervous system to body surfaces, referred to as anterograde axonal trafficking. The virus-encoded protein pUS9 promotes axonal dissemination by sorting virus particles into axons, but whether it is also an effector of fast axonal transport within axons is unknown. To determine the role of pUS9 in anterograde trafficking, we analyzed the axonal transport of pseudorabies virus in the presence and absence of pUS9.

Haemorrhagic Colitis Associated With Enterohaemorrhagic Escherichia Coli O165:H25 Infection In A Yearling Feedlot Heifer, Rodney A. Moxley, Zachary R. Stromberg, Gentry L. Lewis, John D. Loy, Bruce W. Brodersen, Isha R. Patel, Jayanthi Gangiredla

School of Veterinary and Biomedical Sciences: Faculty Publications

Introduction: Enterohaemorrhagic Escherichia coli (EHEC) cause haemorrhagic colitis and haemolytic uraemic syndrome in humans. Although EHEC infection typically results in haemorrhagic colitis in all ages of human patients, in cattle it is usually limited to 1- to 5-week-old nursing calves.

Case Presentation: A 1-year-old feedlot beef heifer was moribund with neurological signs and bloody diarrhoea. At necropsy, the colonic mucosa contained multiple grossly visible haemorrhagic erosions, each measuring <1 mm in diameter. Histologically, foci corresponding to the gross erosions had E. coli O165 antigen-positive bacterial rods adherent to the apical surfaces of degenerate and necrotic colonic mucosal epithelial cells in association with attaching and effacing lesions, and also within cytoplasmic vacuoles in …

Characterization Of Newly Revealed Sequences In The Infectious Myonecrosis Virus Genome In Litopenaeus Vannamei, Duan S. Loy, Sijun Liu, Mark A. Mogler, J. Dustin Loy, Bradley J. Blitvich, Lyric C. Bartholomay

School of Veterinary and Biomedical Sciences: Faculty Publications

Infectious myonecrosis virus (IMNV) causes significant economic losses in farmed shrimp, where associated mortality in ponds can reach 70%. To explore host/pathogen interactions, a next-generation sequencing approach using lymphoid organ tissue from IMNV-infected Litopenaeus vannamei shrimp was conducted. Preliminary sequence assembly of just the virus showed that there were at least an additional 639 bp at the 5′ terminus and 23 nt at the 3′ terminus as compared with the original description of the IMNV genome (7561 nt). Northern blot and reverse transcription-PCR analysis confirmed the presence of novel sequence at both ends of the genome. Using 5′ RACE, an …

Mri-Compatible Bioreactors And Methods Of Using, Shadi Othman, Huihui Xu, Karin Wartella, Vahid Khalilzad-Sharghi, Ian Bargar

School of Veterinary and Biomedical Sciences: Faculty Publications

This disclosure describes a MRI-compatible bioreactor that allows a biological sample to be imaged in culture without disrupting or compromising the culture.

How Do Free-Living, Lerp-Forming, And Gall-Inducing Aphalaridae (Hemiptera: Psylloidea) Affect The Nutritional Quality Of Eucalyptus Leaves?, Anamika Sharma, James Allen, Soundararajan Madhavan, Anantanarayanan Raman, Gary S. Taylor, Murray J. Fletcher

Department of Biochemistry: Faculty Publications

About 380 described species of Psylloidea occur on Eucalyptus in Australia. These show differences in diversification, feeding behavior, and apparent patterns of development. We analyzed the quality of nutrients used by three species of Aphalaridae belonging to different feeding guilds on three species of Eucalyptus. We evaluated the quantity and quality of total nonstructural carbohydrates (TNC), δ¹³C and δ¹⁵N isotope ratios, amino acids, and fatty acids. In general, TNC levels were greater in infested leaves than in uninfested leaves of the three species of Eucalyptus. TNC levels in the leaves of E. macrorhyncha F. …

Overaccumulation Of Γ-Glutamylcysteine In A Jasmonate-Hypersensitive Arabidopsis Mutant Causes Jasmonate-Dependent Growth Inhibition, Hsin-Ho Wei, Martha Rowe, Jean-Jack M. Riethoven, Ryan Grove, Jiri Adamec, Yusuke Jikumaru, Paul E. Staswick

Department of Biochemistry: Faculty Publications

Glutathione (GSH) is essential for many aspects of plant biology and is associated with jasmonate signaling in stress responses. We characterized an Arabidopsis (Arabidopsis thaliana) jasmonate-hypersensitive mutant (jah2) with seedling root growth 100-fold more sensitive to inhibition by the hormone jasmonyl-isoleucine than the wild type. Genetic mapping and genome sequencing determined that the mutation is in intron 6 of GLUTATHIONE SYNTHETASE2, encoding the enzyme that converts γ-glutamylcysteine (γ-EC) to GSH. The level of GSH in jah2 was 71% of the wild type, while the phytoalexin-deficient2-1 (pad2-1) mutant, defective in GSH1 and …

First Evidence For Substrate Channeling Between Proline Catabolic Enzymes A Validation Of Domain Fusion Analysis For Predicting Protein-Protein Interactions, Nikhilesh Sanyal, Benjamin W. Arentson, John J. Tanner, Donald F. Becker

Department of Biochemistry: Faculty Publications

Background: PRODH and P5CDH from Thermus thermophilus are monofunctional enzymes in proline catabolism.

Results: Steady-state kinetics and intermediate trapping data show the PRODH and P5CDH reactions are coupled by a channeling step.

Conclusion: Substrate channeling in monofunctional enzymes is achieved via weak interactions.

Significance: Evidence for substrate channeling between monofunctional proline catabolic enzymes is shown and confirms the Rosetta Stone hypothesis.

Drosophila Muller F Elements Maintain A Distinct Set Of Genomic Properties Over 40 Million Years Of Evolution, Wilson Leung, Cheryl Bailey, Participating Students And Faculty Of The Genomics Education Partnership

Department of Biochemistry: Faculty Publications

The Muller F element (4.2 Mb, ~80 protein-coding genes) is an unusual autosome of Drosophila melanogaster; it is mostly heterochromatic with a low recombination rate. To investigate how these properties impact the evolution of repeats and genes, we manually improved the sequence and annotated the genes on the D. erecta, D. mojavensis, and D. grimshawi F elements and euchromatic domains from the Muller D element. We find that F elements have greater transposon density (25–50%) than euchromatic reference regions (3–11%). Among the F elements, D. grimshawi has the lowest transposon density (particularly DINE-1: 2% vs. 11–27%). F …

Overexpression Of Alpha-Synuclein At Non-Toxic Levels Increases Dopaminergic Cell Death Induced By Copper Exposure Via Modulation Of Protein Degradation Pathways, Annadurai Anandhan, Humberto Rodriguez-Rocha, Iryna Bohovych, Amy M. Griggs, Laura Zavala-Flores, Elsa M. Reyes-Reyes, Javier Seravalli, Lia A. Stanciu, Jaekwon Lee, Jean-Christophe Rochet, Oleh Khalimonchuk, Rodrigo Franco

Department of Biochemistry: Faculty Publications

Gene multiplications or point mutations in alpha (α)-synuclein are associated with familial and sporadic Parkinson’s disease (PD). An increase in copper (Cu) levels has been reported in the cerebrospinal fluid and blood of PD patients, while occupational exposure to Cu has been suggested to augment the risk to develop PD. We aimed to elucidate the mechanisms by which α-synuclein and Cu regulate dopaminergic cell death. Short-term overexpression of WT or A53T α-synuclein had no toxic effect in human dopaminergic cells and primary midbrain cultures, but it exerted a synergistic effect on Cu-induced cell death. Cell death induced by Cu was …

Triacylglycerol Synthesis During Nitrogen Stress Involves The Prokaryotic Lipid Synthesis Pathway And Acyl Chain Remodeling In The Microalgae Coccomyxa Subellipsoidea, James W. Allen, Concetta C. Dirusso, Paul N. Black

Department of Biochemistry: Faculty Publications

Triglyceride (TAG) synthesis during nitrogen starvation and recovery was addressed using Coccomyxa subellipsoidea by analyzing acylchain composition and redistribution using a bioreactor-controlled time course. Galactolipids, phospholipids and TAGs were profiled using liquid chromatography tandem mass spectroscopy (LC–MS/MS). TAG levels increased linearly through 10 days of N starvation to a final concentration of 12.6% dry weight (DW), while chloroplast membrane lipids decreased from 5% to 1.5% DW. The relative quantities of TAG molecular species, differing in acyl chain length and glycerol backbone position, remained unchanged from 3 to 10 days of N starvation. Six TAG species comprised approximately half the TAG …

Metalloprotease Oma1 Fine-Tunes Mitochondrial Bioenergetic Function And Respiratory Supercomplex Stability, Iryna Bohovych, Mario R. Fernandez, Jennifer J. Rahn, Krista D. Stackley, Jennifer E. Bestman, Annadurai Anandhan, Rodrigo Franco, Steven M. Claypool, Robert E. Lewis, Sherine S.L. Chan, Oleh Khalimonchuk

Department of Biochemistry: Faculty Publications

Mitochondria are involved in key cellular functions including energy production, metabolic homeostasis, and apoptosis. Normal mitochondrial function is preserved by several interrelated mechanisms. One mechanism – intramitochondrial quality control (IMQC) – is represented by conserved proteases distributed across mitochondrial compartments. Many aspects and physiological roles of IMQC components remain unclear. Here, we show that the IMQC protease Oma₁ is required for the stability of the respiratory supercomplexes and thus balanced and tunable bioenergetic function. Loss of Oma1 activity leads to a specific destabilization of respiratory supercomplexes and consequently to unbalanced respiration and progressive respiratory decline in yeast. Similarly, experiments …

Rerouting Cellular Electron Flux To Increase The Rate Of Biological Methane Production, Jennie L. Catlett, Alicia M. Ortiz, Nicole R. Baun

Department of Biochemistry: Faculty Publications

Methanogens are anaerobic archaea that grow by producing methane, a gas that is both an efficient renewable fuel and a potent greenhouse gas. We observed that overexpression of the cytoplasmic heterodisulfide reductase enzyme HdrABC increased the rate of methane production from methanol by 30% without affecting the growth rate relative to the parent strain. Hdr enzymes are essential in all known methane-producing archaea. They function as the terminal oxidases in the methanogen electron transport system by reducing the coenzymeM(2-mercaptoethane sulfonate) and coenzyme B (7-mercaptoheptanoylthreonine sulfonate) heterodisulfide, CoM-S-S-CoB, to regenerate the thiol-coenzymes for reuse. In Methanosarcina acetivorans, HdrABC expression caused …

Correction For Sandai Et Al., The Evolutionary Rewiring Of Ubiquitination Targets Has Reprogrammed The Regulation Of Carbon Assimilation In The Pathogenic Yeast Candida Albicans, Doblin Sandai, Zhikang Yin, Laura Selway, David Stead, Janet Walker, Michelle D. Leach, Iryna Bohovych, Iuliana V. Ene, Stavroula Kastora, Susan Budge, Carol A. Munro, Frank C. Odds, Neil A.R. Gow, Alistair J.P. Brown

Department of Biochemistry: Faculty Publications

No abstract provided.

Role Of Micrornas In Alcohol-Induced Multi-Organ Injury, Sathish Kumar Natarajan, Joseph M. Pachunka, Justin L. Mott

Department of Biochemistry: Faculty Publications

Alcohol consumption and its abuse is a major health problem resulting in significant healthcare cost in the United States. Chronic alcoholism results in damage to most of the vital organs in the human body. Among the alcohol-induced injuries, alcoholic liver disease is one of the most prevalent in the United States. Remarkably, ethanol alters expression of a wide variety of microRNAs that can regulate alcohol-induced complications or dysfunctions. In this review, we will discuss the role of microRNAs in alcoholic pancreatitis, alcohol-induced liver damage, intestinal epithelial barrier dysfunction, and brain damage including altered hippocampus structure and function, and neuronal loss, …

Some Of The Most Interesting Casp11 Targets Through The Eyes Of Their Authors, Andriy Kryshtafovych, John Moult, Arnaud Basle, Alex Burgin, Timonthy K. Craig, Robert A. Edwards, Deborah Fass, Marcus D. Hartmann, Mateusz Korycinski, Richard J. Lewis, Donald Lorimer, Andrei N. Lupas, Janet Newman, Thomas S. Peat, Kurt H. Piepenbrink, Janani Prahlad, Mark J. Van Raaij, Forest Rohwer, Anca M. Segall, Victor Seguritan, Eric J. Sundberg, Abhimanyu K. Singh, Mark A. Wilson, Torsten Schwede

Department of Biochemistry: Faculty Publications

The Critical Assessment of protein Structure Prediction (CASP) experiment would not have been possible without the prediction targets provided by the experimental structural biology community. In this article, selected crystallographers providing targets for the CASP11 experiment discuss the functional and biological significance of the target proteins, highlight their most interesting structural features, and assess whether these features were correctly reproduced in the predictions submitted to CASP11.

Organization Of The Mammalian Ionome According To Organ Origin, Lineage Specialization, And Longevity, Siming Ma, Sang-Goo Lee, Eun Bae Kim, Thomas J. Park, Andrei Seluanov, Vera Gorbunova, Rochelle Buffenstein, Javier Seravalli, Vadim N. Gladyshev

Department of Biochemistry: Faculty Publications

Trace elements are essential to all mammals, but their distribution and utilization across species and organs remains unclear. Here, we examined 18 elements in the brain, heart, kidney, and liver of 26 mammalian species and report the elemental composition of these organs, the patterns of utilization across the species, and their correlation with body mass and longevity. Across the organs, we observed distinct distribution patterns for abundant elements, transition metals, and toxic elements. Some elements showed lineage-specific patterns, including reduced selenium utilization in African mole rats, and positive correlation between the number of selenocysteine residues in selenoprotein P and the …

Hare-Mediated Endocytosis Of Hyaluronan And Heparin Is Targeted By Different Subsets Of Three Endocytic Motifs, Madhu S. Pandey, Edward N. Harris, Paul H. Weigel

Department of Biochemistry: Faculty Publications

The hyaluronan (HA) receptor for endocytosis (HARE) is a multifunctional recycling clearance receptor for 14 different ligands, including HA and heparin (Hep), which bind to discrete nonoverlapping sites. Four different functional endocytic motifs (M) in the cytoplasmic domain (CD) target coated pit mediated uptake: (YSYFRI²⁴⁸⁵ (M1), FQHF²⁴⁹⁵ (M2), NPLY²⁵¹⁹ (M3), and DPF²⁵³⁴ (M4)). We previously found (Pandey et al. J. Biol. Chem. 283, 21453, 2008) that M1, M2, and M3 mediate endocytosis of HA. Here we assessed the ability of HARE variants with a single-motif deletion …

Advances In Hyaluronan Biology: Signaling, Regulation, And Disease Mechanisms, Melanie A. Simpson, Carol De La Motte, Larry S. Sherman, Paul H. Weigel

Department of Biochemistry: Faculty Publications

Hyaluronan is an extracellular glycosaminoglycan polymer consisting of linear disaccharide units containing alternating glucuronate and N-acetylglucosamine.Many cell types make hyaluronan, which unlike most other macromolecules is assembled at the plasmamembrane and concurrently translocated through the hyaluronan synthase enzyme. The normal function of large hyaluronan polymers (>1MDa) in tissue cushioning, hydration, and lubrication is well established. The aberrant accumulation and degradation of hyaluronan and the receptor-mediated signaling of smaller hyaluronan fragments have also been extensively implicated in a variety of pathological states including inflammation and cancer. More recently, the discovery that hyaluronan can either be a structural matrix component or …

Salivary Gland Structure Of Ctenarytaina Eucalypti (Maskell, 1890) (Hemiptera) And Phloem Exudate In Eucalyptus Globulus Labillarière, 1799 (Myrtaceae), Anamika Sharma, Soundararajan Madhavan, Anantanarayanan Raman, Gary S. Taylor, Murray J. Fletcher

Department of Biochemistry: Faculty Publications

The structure of the salivary glands of the free-living aphalarid Ctenarytaina eucalypti, which infests multiple species of Eucalyptus in Australasia and has been introduced into many other regions of the world, is described and illustrated. The principal salivary gland is multilobed whereas the accessory gland is tubular. 1-D electrophoresis revealed proteins of approximately 58 and 64 kDa in the salivary gland extracts and proteins of similar molecular weights in the extracted plant exudates, including phloem, from infested leaves and tender shoots of E. globulus. Proteins that could fall within this range include, but are not limited to, …

Janus-Faced Sestrin2 Controls Ros And Mtor Signalling Through Two Separate Functional Domains, Hanseong Kim, Sojin An, Seung-Hyun Ro, Filipa Teixeira, Gyeong Jin Park, Cheal Kim, Chun-Seok Cho, Jeong-Sig Kim, Ursula Jakob, Jun Hee Lee, Uhn-Soo Cho

Department of Biochemistry: Faculty Publications

Sestrins are stress-inducible metabolic regulators with two seemingly unrelated but physiologically important functions: reduction of reactive oxygen species (ROS) and inhibition of the mechanistic target of rapamycin complex 1 (mTORC1). How Sestrins fulfil this dual role has remained elusive so far. Here we report the crystal structure of human Sestrin2 (hSesn2), and show that hSesn2 is twofold pseudo-symmetric with two globular subdomains, which are structurally similar but functionally distinct from each other. While the N-terminal domain (Sesn-A) reduces alkylhydroperoxide radicals through its helix–turn–helix oxidoreductase motif, the C-terminal domain (Sesn-C) modified this motif to accommodate physical interaction with GATOR2 and subsequent …