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Biochemical Journal

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https://www.readbyqxmd.com/read/28935721/the-spliceosomal-proteins-ppih-and-prpf4-exhibit-bipartite-binding
#1
Caroline Rajiv, S RaElle Jackson, Simon Cocklin, Elan Z Eisenmesser, Tara Davis
Pre-mRNA splicing is a dynamic, multi-step process that is catalyzed by the RNA: protein complex called the spliceosome. The spliceosome contains a core set of RNAs and proteins that are conserved in all organisms that perform splicing. In higher organisms, PPIH directly interacts with the core protein PRPF4 and both integrate into the pre-catalytic spliceosome as part of the tri-snRNP sub-complex. As a first step to understand the protein interactions that dictate PPIH and PRPF4 function, we expressed and purified soluble forms of each protein and formed a complex between them...
September 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28935720/suppression-of-store-operated-ca-2-entry-by-activation-of-gper-contribution-to-a-clamping-effect-on-endothelial-ca-2-signaling
#2
Lara E Terry, Mark VerMeer, Jennifer Giles, Quang-Kim Tran
The G Protein-coupled Estrogen Receptor 1 (GPER, formerly a.k.a. GPR30) modulates many Ca(2+)-dependent activities in endothelial cells. However, the underlying mechanisms are poorly understood. We recently reported that GPER acts to prolong cytoplasmic Ca(2+) signals by interacting with and promoting inhibitory phosphorylation of the plasma membrane Ca(2+)-ATPase. In this study, we examined the role of GPER activation in modulating store-operated Ca(2+) entry (SOCE) via effects on the Stromal Interaction Molecule 1 (STIM1)...
September 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28931724/vps18-recruits-vps41-to-the-human-hops-complex-via-a-ring-ring-interaction
#3
Morag Hunter, Edward J Scourfield, Edward Emmott, Stephen C Graham
Eukaryotic cells use conserved multisubunit membrane tethering complexes, including CORVET and HOPS, to control the fusion of endomembranes. These complexes have been extensively studied in yeast, but to date there have been far fewer studies of metazoan CORVET and HOPS. Both of these complexes comprise six subunits: a common four-subunit core and two unique subunits. Once assembled, these complexes function to recognise specific endosomal membrane markers and facilitate SNARE-mediated membrane fusion. CORVET promotes the homotypic fusion of early endosomes, while HOPS promotes the fusion of lysosomes to late endosomes and autophagosomes...
September 20, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28916651/escherichia-coli-and-neisseria-gonorrhoeae-uvrd-helicase-unwinds-g4-dna-structures
#4
Kaustubh Shukla, Roshan Thakur, Debayan Ganguli, Desirazu Rao, Ganesh Nagaraju
G-quadruplex (G4) secondary structures have been implicated in various biological processes including gene expression, DNA replication and telomere maintenance. However, unresolved G4 structures impede replication progression which can lead to generation of DNA double-strand breaks and genome instability. Helicases have been shown to resolve G4 structures to facilitate faithful duplication of the genome. Escherichia coli UvrD (EcUvrD) helicase plays a crucial role in nucleotide excision repair, mismatch repair and in the regulation of homologous recombination...
September 15, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28904078/towards-a-systems-approach-to-human-cytochrome-p450-ensemble-interactions-between-cyp2d6-and-cyp2e1-and-their-functional-consequences
#5
Dmitri R Davydov, Nadezhda Y Davydova, John T Rodgers, Thomas H Rushmore, Jeffrey P Jones
Functional cross-talk among human drug-metabolizing cytochromes P450 through their association is a topic of emerging importance. Here we studied the interactions of human CYP2D6, a major metabolizer of psychoactive drugs, with one of the most prevalent human P450 enzymes, ethanol-inducible CYP2E1. Detection of P450-P450 interactions was accomplished through luminescence resonance energy transfer (LRET) between labeled proteins incorporated into human liver microsomes and the microsomes of insect cells containing NADPH-cytochrome P450 reductase...
September 13, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28899945/bindings-of-no-co-and-o2-to-multi-functional-globin-type-dehaloperoxidase-follow-the-sliding-scale-rule
#6
Gang Wu, Jing Zhao, Stefan Franzen, Ah-Lim Tsai
Dehaloperoxidase-hemoglobin (DHP), a multifunctional globin protein, functions not only as an oxygen carrier as typical globins such as myoglobin, but also as a peroxidase, a mono- and di-oxygenase, peroxygenase, and an oxidase. Kinetics of DHP binding to NO, CO, and O2 were characterized for wild type DHP A and B and the H55D and H55V DHP A mutants. All three gaseous ligands bind to DHP significantly more weakly than sperm whale myoglobin (SWMb). Both CO and NO bind to DHP in a one-step process to form a stable six-coordinate complex...
September 12, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28899944/novel-endogenous-retrovirus-derived-transcript-expressed-in-the-bovine-placenta-is-regulated-by-wnt-signaling
#7
Toshihiro Sakurai, So Nakagawa, Hanako Bai, Rulan Bai, Kazuya Kusama, Atsushi Ideta, Yoshito Aoyagi, Kazuyuki Kaneko, Kosuke Iga, Jiro Yasuda, Takayuki Miyazawa, Kazuhiko Imakawa
Endogenous retroviruses (ERVs) are involved in placentation; perhaps the most well-known ERV s are the syncytins, actively transcribed env genes involved in cell-cell fusion and possible morphological variations. However, ERVs other than syncytins that play an important role in placental development have not been well characterized. To identify ERV genes expressed during the onset of placentation in the bovine species, we characterized the expression profiles of bovine conceptus transcripts during the peri-attachment period using RNA-seq analysis, and confirming some candidates through real-time PCR...
September 12, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28899943/the-chromatin-remodeling-isw1a-complex-is-regulated-by-sumoylation
#8
Qingtang Shen, Nissrine Beyrouthy, Laura Matabishi-Bibi, Catherine Dargemont
The ISWI class of proteins consists in a family of chromatin remodeling ATPases that is ubiquitous in eukaryotes and predominantly functions to slide nucleosomes laterally. The yeast Saccharomyces cerevisiae Isw1 partners with several non-essential alternative subunits-Ioc2, Ioc3, or Ioc4-to form two distinct complexes Isw1a and Isw1b. Besides its ATPase domain, Isw1 presents a C-terminal region formed by HAND, SANT and SLIDE domains responsible for interaction with the Ioc proteins and optimal association of Isw1 to chromatin...
September 12, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28899942/trz1-the-long-form-rnase-z-from-yeast-forms-a-stable-heterohexamer-with-endonuclease-nuc1-and-mutarotase
#9
Miao Ma, Ines Li de La Sierra Gallay, Noureddine Lazar, Olivier Pellegrini, Jean Lepault, Ciarán Condon, Dominique Durand, Herman van Tilbeurgh
Proteomic studies haves established that Trz1, Nuc1 and mutarotase form a complex in yeast. Trz1 is a b-lactamase type RNase composed of two b-lactamase type domains connected by a long linker that is responsible for the endonucleolytic cleavage at the 3'-end of tRNAs during the maturation process (RNase Z activity); Nuc1 is a dimeric mitochondrial nuclease involved in apoptosis, while mutarotase (encoded by YMR099C) catalyzes the conversion between the a- and b-configuration of glucose-6-phosphate. Using gel-filtration, SAXS and electron microscopy we demonstrated that Trz1, Nuc1 and mutarotase form a very stable heterohexamer, composed of two copies of each of the three subunits...
September 12, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28887384/constitutive-activation-of-p46jnk2-is-indispensable-for-c-ebpdelta-induction-in-the-initial-stage-of-adipogenic-differentiation
#10
Joji Kusuyama, Tomokazu Ohnishi, Kenjiro Bandow, Muhammad Subhan Amir, Kaori Shima, Ichiro Semba, Tetsuya Matsuguchi
Adipogenic differentiation plays a vital role in energy homeostasis and endocrine system. Several transcription factors including peroxisome proliferator-activated receptor γ 2 (PPARγ2), CCAAT-enhancer-binding protein (C/EBP) α, β, and δ are important for the process, whereas the stage-specific intracellular signal transduction regulating the onset of adipogenesis remains enigmatic. Here, we explored the functional role of c- jun N-terminal kinases (JNKs) in adipogenic differentiation using in vitro differentiation models of 3T3-L1 cells and primary adipo-progenitor cells...
September 8, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28887383/atspx1-affects-the-atphr1-dna-binding-equilibrium-by-binding-monomeric-atphr1-in-solution
#11
Wanjun Qi, Iain W Manfield, Stephen P Muench, Alison Baker
Phosphorus is an essential macronutrient for plant growth, and is deficient in about 50% of agricultural soils. The transcription factor Phosphate Starvation Response 1 (PHR1) plays a central role in regulating the expression of a subset of Phosphate Starvation Induced (PSI) genes through binding to a cis acting DNA element termed P1BS. In Arabidopsis and rice, activity of AtPHR1/OsPHR2 is regulated in part by their downstream target SPX proteins through protein-protein interaction. Here we provide kinetic and affinity data for interaction between AtPHR1 and P1BS sites...
September 8, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28883123/post-translational-modification-of-the-interferon-gamma-receptor-alters-its-stability-and-signaling
#12
James D Londino, Dexter L Gullick, Travis B Lear, Tomeka L Suber, Nathaniel M Weathington, Luke S Masa, Bill B Chen, Rama K Mallampalli
The IFN gamma receptor 1 (IFNGR1) binds IFN gamma and activates gene transcription pathways crucial for controlling bacterial and viral infections. Although decreases in IFNGR1 surface levels have been demonstrated to inhibit IFN gamma signaling, little is known regarding the molecular mechanisms controlling receptor stability. Here we show in epithelial and monocytic cell lines that IFNGR1 displays K48 polyubiquitination, is proteasomally degraded, and harbors three ubiquitin acceptor sites at K277, K279, and K285...
September 7, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28871039/human-torsina-can-function-in-the-yeast-cytosol-as-a-molecular-chaperone
#13
Ilectra Adam, Lyne Jossé, Mick F Tuite
TorsinA (TorA) is an AAA+ ATPAse linked to dystonia type 1 (DYT1), a neurological disorder that leads to uncontrollable muscular movements. Although DYT1 is linked to a 3bp deletion in the C terminus of TorA, the biological function of TorA remains to be established. Here we use the yeast Saccharomyces cerevisiae as a tractable in vivo model to explore TorA function. We demonstrate that TorA can protect yeast cells against different forms of environmental stress and show that in the absence of the molecular disaggregase Hsp104, TorA can refold heat-denatured luciferase in vivo in an ATP-dependent manner...
September 4, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28864672/the-coproporphyrin-ferrochelatase-of-staphylococcus-aureus-mechanistic-insights-into-a-regulatory-iron-binding-site
#14
Charlie Hobbs, J D Reid, Mark Shepherd
The majority of characterised ferrochelatase enzymes catalyse the final step of classical haem synthesis, inserting ferrous iron into protoporphyrin IX. However, for the recently-discovered coproporphyrin-dependent pathway, ferrochelatase catalyses the penultimate reaction where ferrous iron is inserted into coproporphyrin III. Ferrochelatase enzymes from the bacterial phyla Firmicutes and Actinobacteria have previously been shown to insert iron into coproporphyrin, and those from Bacillus subtilis and Staphylococcus aureus are known to be inhibited by elevated iron concentrations...
September 1, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28864671/expression-of-an-rna-glycosidase-inhibits-hiv-1-transactivation-of-transcription
#15
Meherzad Kutky, Katalin A Hudak
HIV-1 transcription is primarily controlled by the virally encoded Tat protein and its interaction with the viral TAR RNA element. Specifically, binding of a Tat-containing complex to TAR recruits cellular factors that promote elongation of the host RNA polymerase engaging the viral DNA template. Disruption of this interaction halts viral RNA transcription. In this study, we investigated the effect of pokeweed antiviral protein (PAP), an RNA glycosidase (EC#: 3.2.2.22) synthesized by the pokeweed plant ( Phytolacca americana ), on transcription of HIV-1 mRNA...
September 1, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28838949/structural-insights-into-the-substrate-specificity-of-a-glycoside-hydrolase-family-5-lichenase-from-caldicellulosiruptor-sp-f32
#16
Dong-Dong Meng, Xi Liu, Sheng Dong, Ye-Fei Wang, Xiao-Qing Ma, Haixia Zhou, Xinquan Wang, Li-Shan Yao, Yingang Feng, Fu-Li Li
Glycoside hydrolase (GH) family 5 is one of the largest GH families with various GH activities including lichenase, but the structural basis of the GH5 lichenase activity is still unknown. A novel thermostable lichenase F32EG5 belonging to GH5 was identified from an extremely thermophilic bacterium Caldicellulosiruptor  sp. F32. F32EG5 is a bi-functional cellulose and lichenan-degrading enzyme and exhibited a high activity on β-1,3-1,4-glucan but side activity on cellulose. Thin-layer chromatography and NMR analyses indicated that F32EG5 cleaved the β-1,4 linkage or the β-1,3 linkage while a 4- O -substitued glucose residue linked to a glucose residue through a β-1,3 linkage, which is completely different from extensively studied GH16 lichenase that catalyses strict endo-hydrolysis of the β-1,4-glycosidic linkage adjacent to a 3- O -substitued glucose residue in the mixed linked β-glucans...
August 24, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28830911/nitric-oxide-stimulates-cellular-degradation-of-human-cyp51a1-the-highly-conserved-lanosterol-14%C3%AE-demethylase
#17
Ji Won Park, Aria Byrd, Choon-Myung Lee, Edward T Morgan
Nitric oxide (NO) is known to down-regulate drug metabolizing cytochrome P450 enzymes in an enzyme-selective manner. Ubiquitin-proteasome dependent and independent pathways have been reported. Here we studied the regulation of expression of human CYP51A1, the lanosterol 14α-demethylase required for synthesis of cholesterol and other sterols in mammals, which is found in every kingdom of life.  In Huh7 human hepatoma cells, treatment with NO donors caused rapid post-translational down-regulation of CYP51A1 protein...
August 22, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28827282/a-novel-image-based-high-throughput-screening-assay-discovers-therapeutic-candidates-for-adult-polyglucosan-body-disease
#18
Leonardo J Solemsky, Netaly Khazanov, Hanoch Senderowitz, Peixiang Wang, Berge A Minassian, Igor M Ferreira, Wyatt W Yue, Alexander Lossos, Miguel Weil, Or Kakhlon
Glycogen storage disorders (GSDs) are caused by excessive accumulation of glycogen. Some GSDs (Adult Polyglucosan Body Disease (APBD), Tarui and Lafora diseases) are caused by intracellular accumulation of insoluble inclusions, called polyglucosan bodies (PB), which are chiefly composed of malconstructed glycogen. We developed an APBD patient skin fibroblast cell-based assay for PB identification, where the bodies are identified as amylase-resistant periodic acid-Schiff's (PAS) stained structures, and quantified...
August 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28827281/munc18a-clusters-snare-bearing-liposomes-prior-to-trans-snare-zippering
#19
Matthew Grant Arnold, Pratikshya Adhikari, Baobin Kang, Hao Xu
Sec1-Munc18 (SM) proteins cooperate with SNAREs {SNAP [soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein] receptors} to mediate membrane fusion in eukaryotic cells. Studies of Munc18a/Munc18-1/Stxbp1 in neurotransmission suggest that SM proteins accelerate fusion kinetics primarily by activating the partially zippered trans-SNARE complex. However, accumulating evidence has argued for additional roles for SM proteins in earlier steps in the fusion cascade. Here we investigate the function of Munc18a in reconstituted exocytic reactions mediated by neuronal and non-neuronal SNAREs...
August 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28814578/genetic-human-prion-disease-modelled-in-prp-transgenic-drosophila
#20
Alana M Thackray, Alzbeta Cardova, Hanna Wolf, Lydia Pradl, Ina Vorberg, Walker S Jackson, Raymond Bujdoso
Inherited human prion diseases, such as FFI and familial CJD (fCJD), are associated with autosomal dominant mutations in the human prion protein gene PRNP and accumulation of PrPSc, an abnormal isomer of the normal host protein PrPC, in the brain of affected individuals. PrPSc is the principal component of the transmissible neurotoxic prion agent. It is important to identify molecular pathways and cellular processes that regulate prion formation and prion-induced neurotoxicity. This will allow identification of possible therapeutic interventions for individuals with, or at risk from, genetic human prion disease...
August 16, 2017: Biochemical Journal
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