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Effects Of Novel Fragment-Warhead Adducts In Situ And In Vitro With Glutaredoxin Orthologs., Andrew J. Caras 2017 The University of Akron

Effects Of Novel Fragment-Warhead Adducts In Situ And In Vitro With Glutaredoxin Orthologs., Andrew J. Caras

Honors Research Projects

Fragment-based drug design (FBDD) is a method of pharmaceutical research involving characterization of small “fragment” molecule interactions with a target protein. These fragments are optimized with the addition of a nucleophilic “warhead” moiety. Several novel fragment-warhead adducts have been synthesized in house by group member Ram Khattri Ph.D. The experiment investigates reactivity of these adducts with glutaredoxin protein (Grx) orthologs from Homo sapiens, Pseudomonas aeruginosa and Brucella melitensis. Three novel fragment-warhead adducts have been identified to cause weakly bacteriostatic and bacteriocidal effects, most with minimum inhibitory concentration values near 1.28 mg/mL. Discussed herein are molecular effects of ...


Phytochrome B Integrates Light And Temperature Signals In Arabidopsis, Martina Legris, Cornelia Klose, E Sethe Burgie, Cecilia Costigliolo Rojas Rojas, Maximiliano Neme, Andreas Hiltbrunner, Philip A. Wigge, Eberhard Schäfer, Richard D. Vierstra, Jorge J. Casal 2016 Washington University in St Louis

Phytochrome B Integrates Light And Temperature Signals In Arabidopsis, Martina Legris, Cornelia Klose, E Sethe Burgie, Cecilia Costigliolo Rojas Rojas, Maximiliano Neme, Andreas Hiltbrunner, Philip A. Wigge, Eberhard Schäfer, Richard D. Vierstra, Jorge J. Casal

Biology Faculty Publications & Presentations

Ambient temperature regulates many aspects of plant growth and development, but its sensors are unknown. Here, we demonstrate that the phytochrome B (phyB) photoreceptor participates in temperature perception through its temperature-dependent reversion from the active Pfr state to the inactive Pr state. Increased rates of thermal reversion upon exposing Arabidopsis seedlings to warm environments reduce both the abundance of the biologically active Pfr-Pfr dimer pool of phyB and the size of the associated nuclear bodies, even in daylight. Mathematical analysis of stem growth for seedlings expressing wild-type phyB or thermally stable variants under various combinations of light and temperature revealed ...


Expression, Purification, And Crystallization Of An Endoxylanase From Bacteroides Vulgatus, Jesslyn Park, Jason Hurlbert 2016 Winthrop University

Expression, Purification, And Crystallization Of An Endoxylanase From Bacteroides Vulgatus, Jesslyn Park, Jason Hurlbert

The Winthrop McNair Research Bulletin

Sustainable sources of energy are growing in demand as fossil fuels are rapidly expended. One such energy source is fuel ethanol generated from the fermentation of plant biomass by engineered bacterial biocatalysts. The creation of a biocatalyst capable of converting nearly any plant matter to fuel ethanol requires the identification of novel enzymes capable of degrading specific carbohydrate polymers and cloning these enzymes into a bacterial host. This study seeks to structurally characterize a novel xylanase of glycosyl hydrolase family 30 (GH30) from Bacteroides vulgatus, a bacterium found in the human gut microbiome, via x-ray crystallography. The gene for B ...


Heterologous Expression Of A Rice Mir395 Gene In Nicotiana Tabacum Impairs Sulfate Homeostasis, Ning Yuan, Shuangrong Yuan, Zhigang Li, Dayong Li, Qian Hu, Hong Luo 2016 Clemson University

Heterologous Expression Of A Rice Mir395 Gene In Nicotiana Tabacum Impairs Sulfate Homeostasis, Ning Yuan, Shuangrong Yuan, Zhigang Li, Dayong Li, Qian Hu, Hong Luo

Hong Luo

Sulfur participates in many important mechanisms and pathways of plant development. The most common source of sulfur in soil –SO42−– is absorbed into root tissue and distributed into aerial part through vasculature system, where it is reduced into sulfite and finally sulfide within the subcellular organs such as chloroplasts and mitochondria and used for cysteine and methionine biosynthesis. MicroRNAs are involved in many regulation pathways by repressing the expression of their target genes. MiR395 family in Arabidopsis thaliana has been reported to be an important regulator involved in sulfate transport and assimilation, and a high-affinity sulphate transporter and three ATP ...


Ectopic Expression Of A Cyanobacterial Flavodoxin In Creeping Bentgrass Impacts Plant Development And Confers Broad Abiotic Stress Tolerance, Zhigang Li, Shuangrong Yuan, Haiyan Jia, Fangyuan Gao, Qian Hu, Dongfa Sun, Hong Luo 2016 Huazhong Agricultural University

Ectopic Expression Of A Cyanobacterial Flavodoxin In Creeping Bentgrass Impacts Plant Development And Confers Broad Abiotic Stress Tolerance, Zhigang Li, Shuangrong Yuan, Haiyan Jia, Fangyuan Gao, Qian Hu, Dongfa Sun, Hong Luo

Hong Luo

Flavodoxin (Fld) plays a pivotal role in photosynthetic microorganisms as an alternative electron carrier flavoprotein under adverse environmental conditions. Cyanobacterial Fld has been demonstrated to be able to substitute ferredoxin (Fd) of higher plants in most electron transfer processes under stressful conditions. We have explored the potential of Fld for use in improving plant stress response in creeping bentgrass (Agrostis stolonifera L.). Overexpression of Fld altered plant growth and development. Most significantly, transgenic (TG) plants exhibited drastically enhanced performance under oxidative, drought and heat stress as well as nitrogen (N) starvation, which was associated with higher water retention and cell ...


Snp-Based High Density Genetic Map And Mapping Of Btwd1 Dwarfing Gene In Barley, Xifeng Ren, Jibin Wang, Lipan Liu, Genlou Sun, Chengdao Li, Hong Luo, Dongfa Sun 2016 Huazhong Agricultural University

Snp-Based High Density Genetic Map And Mapping Of Btwd1 Dwarfing Gene In Barley, Xifeng Ren, Jibin Wang, Lipan Liu, Genlou Sun, Chengdao Li, Hong Luo, Dongfa Sun

Hong Luo

A high-density linkage map is a valuable tool for functional genomics and breeding. A newly developed sequence-based marker technology, restriction site associated DNA (RAD) sequencing, has been proven to be powerful for the rapid discovery and genotyping of genome-wide single nucleotide polymorphism (SNP) markers and for the high-density genetic map construction. The objective of this research was to construct a high-density genetic map of barley using RAD sequencing. 1894 high-quality SNP markers were developed and mapped onto all seven chromosomes together with 68 SSR markers. These 1962 markers constituted a total genetic length of 1375.8 cM and an average ...


Ataluren Stimulates Ribosomal Selection Of Near-Cognate Trnas To Promote Nonsense Suppression, Bijoyita Roy, Westley J. Friesen, Yuki Tomizawa, John D. Leszyk, Jin Zhuo, Briana Johnson, Jumana Dakka, Christopher R. Trotta, Xiaojiao Xue, Venkateshwar Mutyam, Kim M. Keeling, James A. Mobley, Steven M. Rowe, David M. Bedwell, Ellen M. Welch, Allan Jacobson 2016 University of Massachusetts Medical School

Ataluren Stimulates Ribosomal Selection Of Near-Cognate Trnas To Promote Nonsense Suppression, Bijoyita Roy, Westley J. Friesen, Yuki Tomizawa, John D. Leszyk, Jin Zhuo, Briana Johnson, Jumana Dakka, Christopher R. Trotta, Xiaojiao Xue, Venkateshwar Mutyam, Kim M. Keeling, James A. Mobley, Steven M. Rowe, David M. Bedwell, Ellen M. Welch, Allan Jacobson

Microbiology and Physiological Systems Publications and Presentations

A premature termination codon (PTC) in the ORF of an mRNA generally leads to production of a truncated polypeptide, accelerated degradation of the mRNA, and depression of overall mRNA expression. Accordingly, nonsense mutations cause some of the most severe forms of inherited disorders. The small-molecule drug ataluren promotes therapeutic nonsense suppression and has been thought to mediate the insertion of near-cognate tRNAs at PTCs. However, direct evidence for this activity has been lacking. Here, we expressed multiple nonsense mutation reporters in human cells and yeast and identified the amino acids inserted when a PTC occupies the ribosomal A site in ...


Effects Of Herbicides On Industrial Hemp (Cannabis Sativa) Phytotoxicity, Biomass, And Seed Yield, Brett A. Maxwell 2016 Western Kentucky University

Effects Of Herbicides On Industrial Hemp (Cannabis Sativa) Phytotoxicity, Biomass, And Seed Yield, Brett A. Maxwell

Masters Theses & Specialist Projects

Field studies were established in 2015 at Bowling Green and Lexington, KY to evaluate industrial hemp (Cannabis sativa) tolerance to various herbicides. Hemp was planted into conventionally tilled soils in mid to late June at a seeding rate of 39 kg/ha in Bowling Green and 22 kg/ha in Lexington. Five herbicide active ingredients were applied preemergence (PRE) the day of planting and six postemergence (POST) treatments were applied to 30 cm hemp with a CO2-backpack sprayer delivering 140 L/ha. Plots were 3.1 m wide by 6.1 m long and were sprayed with a 2.1 ...


Therapeutic Targets For Alzheimer's Disease: Insights From In Vitro And In Vivo Models Of Inflammation, Magdalena J. Kiprowska 2016 The Graduate Center, City University of New York

Therapeutic Targets For Alzheimer's Disease: Insights From In Vitro And In Vivo Models Of Inflammation, Magdalena J. Kiprowska

All Graduate Works by Year: Dissertations, Theses, and Capstone Projects

Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive neuronal loss that over the years spreads from the hippocampus to the neural cortex and impairs memory and cognitive functions. At the cellular level AD is linked to the presence of β-amyloid plaques and neurofibrillary tangles but despite decades of research little is known about their contribution to neurodegeneration and whether they are a cause or rather a result of the disease. It is well established that proteasome activity is impaired in AD brains and some studies suggest that this could be one of the initial factors leading to ...


Role Of Hnrnp A1 In The Regulation Of Hdm2 Gene Expression, Heriberto Moran 2016 The Graduate Center, City University of New York

Role Of Hnrnp A1 In The Regulation Of Hdm2 Gene Expression, Heriberto Moran

All Graduate Works by Year: Dissertations, Theses, and Capstone Projects

hnRNP A1 is one of the most abundant and ubiquitously expressed member of hnRNPs (Heterogenous Nuclear Ribonucleoproteins) family of proteins that play multiple roles in gene expression by participating in major steps in the processing of nascent RNA transcripts 1. It is involved in mRNA biogenesis mechanisms such as the transcription, splicing, stability, export and translation of cellular and viral transcripts. The functions of hnRNP A1 extend to the processing of microRNAs, telomere maintenance and DNA repair 1. Our previous studies have shown that hnRNPA1 had reduced protein level and increased cytoplasmic accumulation in senescent human diploid fibroblasts 2,10 ...


Using Mututal Information To Reprogram Dna Specificity Of Laglidadg Endonucleases, Marcon Laforet 2016 The University of Western Ontario

Using Mututal Information To Reprogram Dna Specificity Of Laglidadg Endonucleases, Marcon Laforet

Electronic Thesis and Dissertation Repository

A systematic approach to reprogram protein-DNA interactions has yet to be discovered. This study investigates the ability of co-variation analyses to identify potential protein-DNA contacts that regulate specificity. Here, 27 LAGLIDADG Homing Endonucleases (LHEs) and their 22-basepair DNA targets were collated into a Multiple Sequence Alignment (MSA) that was subjected to pairwise co-variation calculations. Using the LHE I-OnuI as a reference, an amino acid-DNA pair, lysine (K) 231 and adenine +3, generated the highest score. To test if the K231/A3 score was biologically relevant we tested protein mutants for altered nuclease specificity at +3 DNA point mutants. Randomizing the ...


Structural Determinants Allowing Transferase Activity In Sensitive To Freezing 2, Classified As A Family I Glycosyl Hydrolase, Rebecca Roston, Kun Wang, Leslie A. Kuhn, Christoph Benning 2016 University of Nebraska- Lincoln

Structural Determinants Allowing Transferase Activity In Sensitive To Freezing 2, Classified As A Family I Glycosyl Hydrolase, Rebecca Roston, Kun Wang, Leslie A. Kuhn, Christoph Benning

Biochemistry -- Faculty Publications

Background: SENSITIVE TO FREEZING 2 (SFR2) is classified as a glycosyl hydrolase, and by using glycosyltransferase activity, it modifies membrane lipids to promote freeze tolerance.

Results: Although the active site of SFR2 is identical to hydrolases, adjacent loop regions contribute to its transferase activity.

Conclusion: Transferase activity evolved by modifications external to the core catalytic site.

Significance: Defined structure-function relationships will inform engineering of transferases and freeze tolerance.


Ectopic Expression Of A Cyanobacterial Flavodoxin In Creeping Bentgrass Impacts Plant Development And Confers Broad Abiotic Stress Tolerance, Zhigang Li, Shuangrong Yuan, Haiyan Jia, Fangyuan Gao, Qian Hu, Dongfa Sun, Hong Luo 2016 Huazhong Agricultural University

Ectopic Expression Of A Cyanobacterial Flavodoxin In Creeping Bentgrass Impacts Plant Development And Confers Broad Abiotic Stress Tolerance, Zhigang Li, Shuangrong Yuan, Haiyan Jia, Fangyuan Gao, Qian Hu, Dongfa Sun, Hong Luo

Publications

Flavodoxin (Fld) plays a pivotal role in photosynthetic microorganisms as an alternative electron carrier flavoprotein under adverse environmental conditions. Cyanobacterial Fld has been demonstrated to be able to substitute ferredoxin (Fd) of higher plants in most electron transfer processes under stressful conditions. We have explored the potential of Fld for use in improving plant stress response in creeping bentgrass (Agrostis stolonifera L.). Overexpression of Fld altered plant growth and development. Most significantly, transgenic (TG) plants exhibited drastically enhanced performance under oxidative, drought and heat stress as well as nitrogen (N) starvation, which was associated with higher water retention and cell ...


Sending Out An Sos: Mitochondria As A Signaling Hub, Iryna Bohovych, Oleh Khalimonchuk 2016 University of Nebraska- Lincoln

Sending Out An Sos: Mitochondria As A Signaling Hub, Iryna Bohovych, Oleh Khalimonchuk

Biochemistry -- Faculty Publications

Normal cellular physiology is critically dependent on numerous mitochondrial activities including energy conversion, cofactor and precursor metabolite synthesis, and regulation of ion and redox homeostasis. Advances in mitochondrial research during the last two decades provide solid evidence that these organelles are deeply integrated with the rest of the cell and multiple mechanisms are in place to monitor and communicate functional states of mitochondria. In many cases, however, the exact molecular nature of various mitochondria-to-cell communication pathways is only beginning to emerge. Here, we review various signals emitted by distressed or dysfunctional mitochondria and the stress-responsive pathways activated in response to ...


Group I Intron Internal Guide Sequence Binding Strength As A Component Of Ribozyme Network Formation, Elizabeth Satterwhite, Jessica Anne Mellor Yeates, Niles Lehman 2016 Portland State University

Group I Intron Internal Guide Sequence Binding Strength As A Component Of Ribozyme Network Formation, Elizabeth Satterwhite, Jessica Anne Mellor Yeates, Niles Lehman

Chemistry Faculty Publications and Presentations

Origins-of-life research requires searching for a plausible transition from simple chemicals to larger macromolecules that can both hold information and catalyze their own production. We have previously shown that some group I intron ribozymes possess the ability to help synthesize other ribozyme genotypes by recombination reactions in small networks in an autocatalytic fashion. By simplifying these recombination reactions, using fluorescent anisotropy, we quantified the thermodynamic binding strength between two nucleotides of two group I intron RNA fragments for all 16 possible genotype combinations. We provide evidence that the binding strength (KD) between the 3-nucleotide internal guide sequence (IGS) of ...


Topographical Expression Of Class Ia And Class Ii Phosphoinositide 3-Kinase Enzymes In Normal Human Tissues Is Consistent With A Role In Differentiation, Soha Salama El Sheikh, Jan Domin, Prakitpunthu Tomtitchong, Paul Abel, Gordon Stamp, El-Nasir Lalani 2016 Imperial College School of Medicine

Topographical Expression Of Class Ia And Class Ii Phosphoinositide 3-Kinase Enzymes In Normal Human Tissues Is Consistent With A Role In Differentiation, Soha Salama El Sheikh, Jan Domin, Prakitpunthu Tomtitchong, Paul Abel, Gordon Stamp, El-Nasir Lalani

El Nasir Lalani

Background: Growth factor, cytokine and chemokine-induced activation of PI3K enzymes constitutes the start of a complex signalling cascade, which ultimately mediates cellular activities such as proliferation, differentiation, chemotaxis, survival, trafficking, and glucose homeostasis. The PI3K enzyme family is divided into 3 classes; class I (subdivided into IA and IB), class II (PI3K-C2α, PI3K-C2β and PI3K-C2γ) and class III PI3K. Expression of these enzymes in human tissue has not been clearly defined.

Methods: In this study, we analysed the immunohistochemical topographical expression profile of class IA (anti-p85 adaptor) and class II PI3K (PI3K-C2α and PI3K-C2β) enzymes in 104 formalin-fixed, paraffin embedded ...


Suppression Of Ischemia In Arterial Occlusive Disease By Jnk-Promoted Native Collateral Artery Development, Kasmir Ramo, Koichi Sugamura, Siobhan M. Craige, John F. Keaney Jr., Roger J. Davis 2016 University of Massachusetts Medical School

Suppression Of Ischemia In Arterial Occlusive Disease By Jnk-Promoted Native Collateral Artery Development, Kasmir Ramo, Koichi Sugamura, Siobhan M. Craige, John F. Keaney Jr., Roger J. Davis

Davis Lab

Arterial occlusive diseases are major causes of morbidity and mortality. Blood flow to the affected tissue must be restored quickly if viability and function are to be preserved. We report that disruption of the mixed-lineage protein kinase (MLK) - cJun NH2-terminal kinase (JNK) signaling pathway in endothelial cells causes severe blockade of blood flow and failure to recover in the murine femoral artery ligation model of hindlimb ischemia. We show that the MLK-JNK pathway is required for the formation of native collateral arteries that can restore circulation following arterial occlusion. Disruption of the MLK-JNK pathway causes decreased Dll4/Notch signaling, excessive ...


Autophagic Turnover Of Inactive 26s Proteasomes In Yeast Is Directed By The Ubiquitin Receptor Cue5 And The Hsp42 Chaperone, Richard S. Marshall, Fionn McLoughlin, Richard D. Vierstra 2016 Washington University in St Louis

Autophagic Turnover Of Inactive 26s Proteasomes In Yeast Is Directed By The Ubiquitin Receptor Cue5 And The Hsp42 Chaperone, Richard S. Marshall, Fionn Mcloughlin, Richard D. Vierstra

Biology Faculty Publications & Presentations

Highlights

  • The yeast 26S proteasome is degraded by Atg8-mediated autophagy
  • Nitrogen starvation and inactivation stimulate proteaphagy via distinct pathways
  • Proteasome inhibition is accompanied by extensive ubiquitylation of the complex
  • Proteaphagy engages the Cue5 autophagy receptor and the Hsp42 chaperone

Summary

The autophagic clearance of 26S proteasomes (proteaphagy) is an important homeostatic mechanism within the ubiquitin system that modulates proteolytic capacity and eliminates damaged particles. Here, we define two proteaphagy routes in yeast that respond to either nitrogen starvation or particle inactivation. Whereas the core autophagic machineries required for Atg8 lipidation and vesiculation are essential for both routes, the upstream Atg1 ...


Expression And Site-Directed Mutagenesis Of Type Iii Polyketide Synthases, Logan R. Richards, Odessa Goudy, Lee M. Stunkard, Aaron B. Benjamin, Bethany P. Manning, Jeremy Lohman 2016 Purdue University

Expression And Site-Directed Mutagenesis Of Type Iii Polyketide Synthases, Logan R. Richards, Odessa Goudy, Lee M. Stunkard, Aaron B. Benjamin, Bethany P. Manning, Jeremy Lohman

The Summer Undergraduate Research Fellowship (SURF) Symposium

Natural products are a well-established source of drugs, and evolution has yielded polyketides such as leinamycin and iso-migrastatin that have demonstrated anti-tumor activity. Polyketides are large metabolites with a high degree of chemical variability and are commonly produced by soil bacteria. Polyketide synthases (PKS) exist as three different archetypes, and the reaction mechanisms of ketosynthases from all archetypes is not understood. Type III PKSs exist as an independently functioning ketosynthase (KS), which primarily use coenzyme A (CoA), with some exceptions, for the biosynthesis of polyketides. We elected to focus our studies on ketosynthases, because they are responsible for forming the ...


Role Of Sumoylation In Mitochondrial Division In Tetrahymena Thermophila, Ramya Modi, James Forney 2016 Purdue University

Role Of Sumoylation In Mitochondrial Division In Tetrahymena Thermophila, Ramya Modi, James Forney

The Summer Undergraduate Research Fellowship (SURF) Symposium

SUMOylation is a post translation modification that involves the addition of a small protein called SUMO, Small Ubiquitin-like MOdifier to a target protein. It is an important mechanism for the regulation of gene expression, the maintenance of genomic stability and in modifying nuclear proteins. More recently evidence has emerged for its importance in regulating mitochondrial fission and fusion in mammalian cells. This study evaluates the parameters for optimal staining of Tetrahymena thermophila mitochondria using two different dyes and then examines different cell lines with defects in the SUMOylation pathway. The first staining method uses Mitotracker Green, a vital stain that ...


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