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Ginger D Blonde, Susan P Travers, Alan Craig Spector
The taste of L-glutamate and its synergism with 5'-ribonucleotides is thought to be primarily mediated through the T1R1+T1R3 heterodimer in some mammals including rodents and humans. While knockout (KO) mice lacking either receptor subunit show impaired sensitivity to a range of monosodium glutamate (MSG) concentrations mixed with 2.5mM inosine 5'-monophosphate (IMP) in amiloride, wild-type (WT) controls can detect this IMP concentration, hindering direct comparison between genotypes. Moreover, some residual sensitivity persists in the KO group, suggesting that the remaining subunit could maintain a limited degree of function...
February 14, 2018: American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
Emily L Gulliver, Amy Wright, Deanna Deveson Lucas, Marianne Mégroz, Oded Kleifeld, Ralf Schittenhelm, David Powell, Torsten Seemann, Juergen Bulitta, Marina Harper, John D Boyce
Pasteurella multocida is a Gram-negative bacterium responsible for many important animal diseases. While a number of P. multocida virulence factors have been identified, very little is known about how gene expression and protein production is regulated in this organism. Small RNA (sRNA) molecules are critical regulators that act by binding to specific mRNA targets, often in association with the RNA chaperone protein Hfq. In this study, transcriptomic analysis of the P. multocida strain VP161 revealed a putative sRNA with high identity to GcvB from Escherichia coli and Salmonella enterica serovar Typhimurium...
February 12, 2018: RNA
Maria Oszvald, Lucia F Primavesi, Cara A Griffiths, Jonathan Cohn, Shib Basu, Michael L Nuccio, Matthew J Paul
Transgenic maize expressing a rice trehalose phosphate phosphatase (TPP1) controlled by a rice MADS6 promoter active over the 2- to 3-week flowering period are higher yielding than wild type with or without drought during flowering. To better understand the mechanistic basis of the yield increase, we characterise gene expression and metabolite profiles in developing female reproductive tissue comprising florets, node, pith and shank and in leaves over the flowering period with and without drought. Using the β-glucuronidase reporter the MADS6 promoter activity was most closely linked to vasculature, particularly phloem companion cells in florets and pith consistent with largest decreases in trehalose 6-phosphate (T6P) levels (2-3 fold) in pith and florets...
February 6, 2018: Plant Physiology
Olatz Villate, Nekane Ibarluzea, Eugenia Fraile-Bethencourt, Alberto Valenzuela, Eladio A Velasco, Detelina Grozeva, F L Raymond, María P Botella, María-Isabel Tejada
Mutations in CHD7 have been shown to be a major cause of CHARGE syndrome, which presents many symptoms and features common to other syndromes making its diagnosis difficult. Next generation sequencing (NGS) of a panel of intellectual disability related genes was performed in an adult patient without molecular diagnosis. A splice donor variant in CHD7 (c.5665 + 1G > T) was identified. To study its potential pathogenicity, exons and flanking intronic sequences were amplified from patient DNA and cloned into the pSAD® splicing vector...
2018: Frontiers in Genetics
Isabel Egea, Irene Albaladejo, Victoriano Meco, Belén Morales, Angel Sevilla, Maria C Bolarin, Francisco B Flores
Breeding for drought-tolerant crops is a pressing issue due to the increasing frequency and duration of droughts caused by climate change. Although important sources of variation for drought tolerance exist in wild relatives, the mechanisms and the key genes controlling tolerance in tomato are little known. The aim of this study is to determine the drought response of the tomato wild relative Solanum pennellii (Sp) compared with the cultivated tomato Solanum lycopersicum (Sl). The paper investigates the physiological and molecular responses in leaves of Sp and Sl plants without stress and moderate drought stress...
February 12, 2018: Scientific Reports
Nikolay A Alemasov, Nikita V Ivanisenko, Srinivasan Ramachandran, Vladimir A Ivanisenko
BACKGROUND: So far, little is known about the molecular mechanisms of amyotrophic lateral sclerosis onset and progression caused by SOD1 mutations. One of the hypotheses is based on SOD1 misfolding resulting from mutations and subsequent deposition of its cytotoxic aggregates. This hypothesis is complicated by the fact that known SOD1 mutations of similar clinical effect could be distributed over the whole protein structure. RESULTS: In this work, a measure of hydrogen bond stability in conformational states was studied with elastic network analysis of 35 SOD1 mutants...
February 5, 2018: BMC Structural Biology
Jia Ying Cheang, Peter Michael Moyle
Glucagon-like peptide-1 (GLP-1) is secreted by intestinal L-cells following food intake, and plays an important role in glucose homeostasis due to its stimulation of glucose-dependent insulin secretion. Further, GLP-1 is also associated with protective effects on pancreatic β-cells and the cardiovascular system, reduced appetite, and weight loss, making GLP-1 derivatives an exciting treatment for type 2 diabetes and obesity. Despite these benefits, wild-type GLP-1 exhibits a short circulation time, due to its poor metabolic stability and rapid renal clearance, and needs to be administered by injection, making it a poor therapeutic agent...
February 11, 2018: ChemMedChem
Neta Luria, Elisheva Smith, Noa Sela, Oded Lachman, Ilana Bekelman, Amnon Koren, Aviv Dombrovsky
During October 2014, unfamiliar mild mosaic and mottling symptoms were identified on leaves of pepper (Capsicum chinense cv. Habanero) seedlings grown in the Arava valley in Israel 2-3 weeks post planting. Symptomatic plants were tested positive by ELISA using laboratory-produced antisera for tobamovirus species. Typical tobamovirus rod-shaped morphology was observed by transmission electron microscopy (TEM) analysis of purified virion preparation that was used for mechanical inoculation of laboratory test plants for the completion of Koch's postulates...
February 10, 2018: Virus Genes
Huanying Pang, Mingsheng Qiu, Jingmin Zhao, Rowena Hoare, Sean J Monaghan, Dawei Song, Yunsheng Chang, Jichang Jian
Vibrio alginolyticus, a bacterial pathogen in fish and humans, expresses a type III secretion system (T3SS) that is critical for pathogen virulence and disease development. However, little is known about the associated effectors (T3SEs) and their physiological role. In this study, the T3SE gene hopPmaJ (hop) was cloned from V. alginolyticus wild-type strain HY9901 and the mutant strain HY9901Δhop was constructed by the in-frame deletion method. The results showed that the deduced amino acid sequence of V. alginolyticus HopPmaJ shared 78-98% homology with other Vibrio spp...
February 7, 2018: Fish & Shellfish Immunology
Kiran Kurmi, Sadae Hitosugi, Elizabeth K Wiese, Felix Boakye-Agyeman, Wilson I Gonsalves, Zhenkun Lou, Larry M Karnitz, Matthew P Goetz, Taro Hitosugi
Lysine succinylation was recently identified as a post-translational modification in cells. However, the molecular mechanism underlying lysine succinylation remains unclear. Here, we show that carnitine palmitoyltransferase 1A (CPT1A) has lysine succinyltransferase (LSTase) activity in vivo and in vitro. Using a stable isotope labeling by amino acid in cell culture (SILAC)-based proteomics approach, we found that 101 proteins were more succinylated in cells expressing wild-type (WT) CPT1A compared with vector control cells...
February 6, 2018: Cell Reports
Qian Yang, Wei Deng, Shipeng Wang, Hongjie Liu, Xuefeng Li, Mingqi Zheng
BACKGROUND: Descurainia sophia L., a problematic weed in winter wheat fields in China, has developed high resistance to tribenuron-methyl. Amino acid substitutions at sites Pro197, Asp376 and Trp574 in the target acetohydroxyacid synthase (AHAS) were primarily responsible for D. sophia resistance to tribenuron-methyl. In this study, D. sophia plants individually homozygous for a specific resistance mutation (Pro197Leu, Pro197His, Pro197Ser, Pro197Thr, Asp376Glu or Trp574Leu) in AHAS were purified and generated, and the effects of resistance mutations on D...
February 9, 2018: Pest Management Science
Lei Chen, Sheng Chen, Jing Wu, Dan Wu
We used in vitro molecular evolution technology by error-prone PCR and high-throughput screening to improve thermostability of Bacillus flexus CCTCC 2015368 β-amylase. Mutant D476N with significant thermostability increase was selected by LB agar starch plate colorimetric assay and 96-well plate enzyme activity assay. The optimum pH was 6.5 for the mutant D476N, compared to 7.0 of the wild type. The optimal temperature was 55 ℃ for both mutant D476N and the wild type. The T₅₀ value of the mutant D476N was 4 ℃ higher than that of the wild type...
February 25, 2018: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Dianjun Cao, Yan-Yan Ni, Xiang-Jin Meng
BACKGROUND: Genotype 3 hepatitis E virus (HEV) infection is generally associated with mild disease. However, recently eight genotype 3 HEV isolates were identified from patients with severe hepatitis. Importantly, three mutations (S605P, I978V and V1213A) in these genotype 3 isolates were found to be typical of genotype 4 HEV, which is sometime associated with more severe hepatitis. Therefore in this study we seek to determine if these unique mutations contribute to enhanced virus replication and thus potentially severe disease...
February 8, 2018: Virology Journal
Muhammad Irfan, Claudio F Gonzalez, Saad Raza, Muhamamd Rafiq, Fariha Hasan, Samiullah Khan, Aamer Ali Shah
Enzymes activity and stability at extreme temperature can be intensified by regularly applying protein engineering. In the present study, two amino acids were perceived to mark the temperature dependability of xylanase from Geobacillus thermodenitrificans C5. Six mutants of G. thermodenitrificans C5 were built through site-directed mutagenesis by interchanging the residue with proline and glutamic acid (R81P, H82E, W185P, D186E, double mutant W185P/D186E and triple mutant H82E/W185P/D186E). Both mutant and wild type enzymes were quantified in host E...
April 2018: Enzyme and Microbial Technology
William E Plautz, Vijaya Satish Sekhar Pilli, Brian C Cooley, Rima Chattopadhyay, Pamela R Westmark, Todd Getz, David Paul, Wolfgang Bergmeier, John P Sheehan, Rinku Majumder
OBJECTIVE: PS (protein S) is a plasma protein that directly inhibits the coagulation FIXa (factor IXa) in vitro. Because elevated FIXa is associated with increased risk of venous thromboembolism, it is important to establish how PS inhibits FIXa function in vivo. The goal of this study is to confirm direct binding of PS with FIXa in vivo, identify FIXa amino acid residues required for binding PS in vivo, and use an enzymatically active FIXa mutant that is unable to bind PS to measure the significance of PS-FIXa interaction in hemostasis...
February 1, 2018: Arteriosclerosis, Thrombosis, and Vascular Biology
Liping Ma, Tao Jin, Hanzhong Wang, Haizhou Liu, Runkun Wang, Yong Li, Guoxiang Yang, Yanping Xiong, Jing Chen, Jun Zhang, Guang Chen, Wei Li, Di Liu, Peng Lin, Yueying Huang, George F Gao, Quanjiao Chen
Since 2016, the highly pathogenic avian influenza H5N8 virus has emerged in the Central Asian flyway and Europe, causing massive deaths in poultry and wild birds. In this study, we isolated and identified three H5N8 viruses from swan goose and black swans in Hubei province during the 2016/2017 winter season. Whole-genome sequencing and phylogenetic analysis revealed that the three viruses clustered into a group of H5N8 viruses from Qinghai Lake and Europe. A novel reassortment virus from swan goose was distinguished from that of black swans, in that its PA and NP genes were distinct from those of Qinghai Lake viruses...
February 7, 2018: Emerging Microbes & Infections
M Giovanola, A Vollero, R Cinquetti, E Bossi, L R Forrest, E Di Cairano, M Castagna
The crystallizations of the prokaryotic LeuT and of the eukaryotic DAT and SERT transporters represent important steps forward in the comprehension of the molecular physiology of Neurotransmitter:Sodium Symporters, although the molecular determinants of the coupling mechanism and of ion selectivity still remain to be fully elucidated. The insect NSS homologue KAAT1 exhibits unusual physiological features, such as the ability to use K+ as the driver ion, weak chloride dependence, and the ability of the driver ion to influence the substrate selectivity; these characteristics can help to define the molecular determinants of NSS function...
January 30, 2018: Biochimica et Biophysica Acta
Cinzia Pisani, Georgios Strimpakos, Francesca Gabanella, Maria Grazia Di Certo, Annalisa Onori, Cinzia Severini, Siro Luvisetto, Stefano Farioli-Vecchioli, Irene Carrozzo, Antonio Esposito, Tamara Canu, Elisabetta Mattei, Nicoletta Corbi, Claudio Passananti
Up-regulation of the dystrophin-related gene utrophin represents a promising therapeutic strategy for the treatment of Duchenne Muscular Dystrophy (DMD). In order to re-program the utrophin expression level in muscle, we engineered artificial zinc finger transcription factors (ZF-ATFs) that target the utrophin "A" promoter. We have previously shown that the ZF-ATF "Jazz", either by transgenic manipulation or by systemic adeno-associated viral delivery, induces significant rescue of muscle function in dystrophic "mdx" mice...
January 30, 2018: Biochimica et Biophysica Acta
Chong Zuo, Amber L Jolly, Andriana P Nikolova, David I Satzer, Sirui Cao, Jeremy S Sanchez, David P Ballou, Elizabeth E Trimmer
The flavoprotein methylenetetrahydrofolate reductase (MTHFR) from Escherichia coli catalyzes a ping-pong reaction with NADH and 5,10-methylenetetrahydrofolate (CH2-H4folate) to produce NAD+ and 5-methyltetrahydrofolate (CH3-H4folate). This work focuses on the function of the invariant, active-site amino acid Gln183. X-ray structures of the enzyme complexes Ered(wild-type)•NADH and Eox(Glu28Gln)•CH3-H4folate indicate that Gln183 makes key hydrogen-bonding interactions with both NADH and folate in their respective half-reactions, suggesting roles in binding each substrate...
February 3, 2018: Archives of Biochemistry and Biophysics
Cynthia K Hollland, Daniel A Berkovich, Madeleine L Kohn, Hiroshi Maeda, Joseph M Jez
Aromatic amino acids are protein building blocks and precursors to a number of plant natural products, such as the structural polymer lignin and a variety of medicinally relevant compounds. Plants make tyrosine and phenylalanine by a different pathway from many microbes, which requires prephenate aminotransferase (PAT) as the key enzyme. PAT produces arogenate, the unique and immediate precursor for both tyrosine and phenylalanine in plants, and also has aspartate aminotransferase (AAT) activity. The molecular mechanisms governing the substrate specificity and activation or inhibition of PAT are currently unknown...
February 5, 2018: Plant Journal: for Cell and Molecular Biology
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