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Journal of Biological Chemistry

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https://www.readbyqxmd.com/read/28104805/microrna-145-modulates-n6-methyladenosine-levels-by-targeting-the-3-untranslated-mrna-region-of-the-n6-methyladenosine-binding-yth-domain-family-2-protein
#1
Zhe Yang, Jiong Li, Guoxing Feng, Shan Gao, Yuan Wang, Shuqin Zhang, Yunxia Liu, Lihong Ye, Yueguo Li, Xiaodong Zhang
N6 methyladenosine (m6A) is a prevalent modification present in the mRNAs of higher eukaryotes. YTH domain family 2 (YTHDF2), an m6A reader protein, can recognize mRNA m6A sites to mediate mRNA degradation. However, the regulatory mechanism of YTHDF2 is poorly understood. To this end, we investigated the post transcriptional regulation of YTHDF2. Bioinformatics analysis suggested that the microRNA miR 145 might target the 3'-untranslated region (3'UTR) of YTHDF2 mRNA. The levels of miR 145 were negatively correlated with those of YTHDF2 mRNA in clinical hepatocellular carcinoma (HCC) tissues, and immunohistochemical staining revealed that YTHDF2 was closely associated with malignancy of HCC...
January 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28104804/the-n-terminus-specifies-the-switch-between-transport-modes-of-the-human-serotonin-transporter
#2
Carina Kern, Fatma Asli Erdem, Ali El-Kasaby, Walter Sandtner, Michael Freissmuth, Sonja Sucic
The serotonin transporter (SERT) and other monoamine transporters operate in either a forward transport mode, where the transporter undergoes a full transport cycle, or an exchange mode, where the transporter seesaws through half-cycles. Amphetamines trigger the exchange-mode leading to substrate efflux. This efflux was proposed to rely on the N-terminus, which was suggested to adopt different conformations in the inward-, outward-facing and amphetamine-bound states. This prediction was verified by tryptic digestion of SERT-expressing membranes: in the absence of Na+, the N-terminus was rapidly digested...
January 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28104803/microrna-processing-enzymes-are-essential-for-survival-and-function-of-mature-retinal-pigmented-epithelial-cells-in-mice
#3
Thomas R Sundermeier, Sanae Sakami, Bhubanananda Sahu, Scott J Howell, Songqi Gao, Zhiqian Dong, Marcin Golczak, Akiko Maeda, Krzysztof Palczewski
Age-related macular degeneration (AMD) is a major cause of irreversible vision loss. The neovascular or "wet" form of AMD can be treated to a varying degree with anti-angiogenic drugs, but geographic atrophy (GA) is an advanced stage of the more prevalent "dry" form of AMD for which there is no effective treatment. Development of GA has been linked to loss of the microRNA (miRNA)-processing enzyme DICER1 in the mature retinal pigmented epithelium (RPE). This loss results in the accumulation of toxic transcripts of Alu transposable elements which activate the NLRP3 inflammasome and additional downstream pathways that compromise the integrity and function of the RPE...
January 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28104802/the-ssl2245-sll1130-toxin-antitoxin-system-mediates-heat-induced-programmed-cell-death-in-synechocystis-sp-pcc6803
#4
Afshan Srikumar, Pilla Sankara Krishna, Dokku Sivaramakrishna, Stefan Kopfmann, Wolfgang R Hess, Musti J Swamy, Sue Lin-Chao, Jogadhenu S S Prakash
Two putative heat responsive genes, ssl2245 and sll1130 constitute an operon that also has characteristics of a toxin-antitoxin system, thus joining several enigmatic features. Closely related orthologs of Ssl2245 and Sll1130 exist in widely different bacteria, which thrive under environments with large fluctuations in temperature and salinity, among which some are thermo-epilithic biofilm forming cyanobacteria. Transcriptome analyses revealed that the CRISPR genes as well as several hypothetical genes were commonly up-regulated in Δssl2245 and Δsll1130 mutants...
January 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100787/polymorphic-variants-of-human-protein-l-isoaspartyl-methyltransferase-affect-catalytic-activity-aggregation-and-thermal-stability-implications-for-the-etiology-of-neurological-disorders-and-cognitive-aging
#5
Charity Juang, Baihe Chen, Jean-Louis Bru, Katherine Nguyen, Eric Huynh, Mahsa Momen, Jeungjin Kim, Dana W Aswad
Protein L-isoaspartyl methyltransferase (PIMT/PCMT1), a product of the human pcmt1 gene, catalyzes repair of abnormal L-isoaspartyl linkages in age-damaged proteins. Pcmt1 knockout mice exhibit a profound neuropathology and die 30-60 days postnatal from an epileptic seizure. Here we express 15 reported variants of human PIMT and characterize them with regard to their enzymatic activity, thermal stability, and propensity to aggregation. One mutation, R36C, renders PIMT completely inactive, while two others, A7P and I58V, exhibit activity that is 80-100% higher than wild type...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100786/inositol-depletion-induced-by-acute-treatment-of-the-bipolar-disorder-drug-valproate-increases-levels-of-phytosphingosine
#6
Shyamalagauri Jadhav, Sarah Russo, L Ashley Cowart, Miriam L Greenberg
Bipolar disorder (BD) is a severe psychiatric illness affecting ~2% of the world population. Valproate (VPA) and lithium, though widely used for the treatment of BD, are not universally effective. These drugs have been shown to cause inositol depletion, but translating this observation to a specific therapeutic mechanism has been difficult, hampering the development of more effective therapies. We have shown previously in yeast that chronic VPA treatment induces the unfolded protein response due to increasing ceramide levels...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100785/protein-kinase-c-controls-binding-of-igo-ensa-proteins-to-protein-phosphatase-2a-in-budding-yeast
#7
Vu Thai, Noah Dephoure, Amit Weiss, Jacqueline Ferguson, Ricardo Leitao, Steven P Gygi, Douglas R Kellogg
Protein phosphatase 2A (PP2A) plays important roles in controlling mitosis in all eukaryotic cells. The form of PP2A that controls mitosis is associated with a conserved regulatory subunit that is called B55 in vertebrates and Cdc55 in budding yeast. The activity of this form of PP2A can be inhibited by binding of conserved Igo/ENSA proteins. Although the mechanisms that activate Igo/ENSA to bind to PP2A are well understood, little is known about how they are inactivated. Here, we have analyzed regulation of Igo/ENSA in the context of a checkpoint pathway that links mitotic entry to membrane growth in budding yeast...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100784/mitochondrial-o-glcnac-transferase-mogt-regulates-mitochondrial-structure-function-and-survival-in-hela-cells
#8
Juliana L Sacoman, Raul Y Dagda, Amanda R Burnham-Marusich, Ruben K Dagda, Patricia M Berninsone
O-linked N-acetylglucosamine transferase (OGT) catalyzes O-GlcNAcylation of target proteins and regulates numerous biological processes. OGT is encoded by a single gene that yields nucleocytosolic and mitochondrial isoforms. To date, the role of the mitochondrial isoform of OGT (mOGT) remains largely unknown. Using high-throughput proteomics, we identified 84 candidate mitochondrial glycoproteins, of which 44 are novel. Notably, two of the candidate glycoproteins identified (cytochrome oxidase 2 (COX2) and NADH: ubiquinone oxidoreductase core subunit 4 (MT-ND4)) are encoded by mitochondrial DNA...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100783/dual-functional-roles-of-molecular-beacon-a-microrna-detector-and-inhibitor
#9
Wai Ming Li, Ching-Man Chan, Andrew L Miller, Chow H Lee
MicroRNAs are essential in many cellular processes. The ability to detect microRNAs is important for understanding its function and biogenesis. This study is aimed at using a molecular beacon to detect miR-430 in developing zebrafish embryos as a proof of principle. miR-430 is crucial for the clearance of maternal mRNA during maternal zygotic transition (MZT) in embryonic development. Despite its known function, the temporal and spatial expression of miR-430 remains unclear. We used various imaging techniques, including laser scanning confocal microscopy, spinning disk and lightsheet microscopy, to study the localization of miR-430 and any developmental defects possibly caused by the molecular beacon...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100782/triphosphate-re-orientation-of-the-incoming-nucleotide-as-a-fidelity-checkpoint-in-viral-rna-dependent-rna-polymerases
#10
Xiaorong Yang, Xinran Liu, Derek M Musser, Ibrahim M Moustafa, Jamie J Arnold, Craig E Cameron, David D Boehr
The nucleotide incorporation fidelity of the viral RNA-dependent RNA polymerase (RdRp) is important for maintaining functional genetic information, but at the same time for generating sufficient genetic diversity to escape the bottlenecks of the host's anti-viral response. We have previously shown that the structural dynamics of the motif-D loop is closely related to nucleotide discrimination. Previous studies have also suggested that there is a re-orientation of the triphosphate of the incoming nucleotide, which is essential prior to nucleophilic attack from the primer RNA 3'-hydroxyl...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100781/biogenesis-of-a-mitochondrial-outer-membrane-protein-in-trypanosoma-brucei-targeting-signal-and-dependence-on-an-unique-biogenesis-factor
#11
Julia Bruggisser, Sandro K Aumlser, Jan Mani, Andre Schneider
The mitochondrial outer membrane (OM) contains single and multiple membrane-spanning proteins which need to contain signals that ensure correct targeting and insertion into the OM. The biogenesis of such proteins has so far essentially only been studied in yeast and related organisms. Here we show that POMP10, an OM protein of the early diverging protozoan Trypanosoma brucei, is signal-anchored. Transgenic cells expressing variants of POMP10 fused to GFP demonstrate that the N-terminal membrane-spanning domain flanked by a few positively charged or neutral residues is both necessary and sufficient for mitochondrial targeting...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100780/a-novel-microrna-located-in-trkc-gene-regulates-wnt-signaling-pathway-and-is-differentially-expressed-in-colorectal-cancer-specimens
#12
Sadat Dokanehiifard, Atena Yasari, Hadi Najafi, Meisam Jafarzadeh, Maryam Nikkhah, Seyed Javad Mowla, Bahram M Soltani
Tropomyosin receptor kinase C (TrkC) is involved in cell survival, apoptosis, differentiation and tumorigenesis. TrkC diverse functions might be attributed to the hypothetical non-coding RNAs, embedded within the gene. Using bioinformatics approaches, a novel miRNA named TrkC-miR2 was predicted within TrkC gene, capable of regulating Wnt pathway. For experimental verification of this miRNA, predicted TrkC-premir2 sequence was overexpressed in SW480 cells, which led to the detection of two mature TrkC-miR2 isomiRs, and their endogenous forms were detected in human cell lines as well...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100779/intramembrane-proteolysis-of-astrotactins
#13
Hao Chang, Philip M Smallwood, John Williams, Jeremy Nathans
Astrotactins are vertebrate-specific membrane proteins implicated in neuron-glia interactions during central nervous system development and in hair follicle polarity during skin development. By studying epitope-tagged derivatives of mouse Astrotactin2 (Astn2) produced in transfected cells, we determined that the amino- and carboxy-termini reside in the extracellular space and are initially linked by two transmembrane segments and a single cytoplasmic domain. We further show that Astn2 undergoes proteolytic cleavage in the second transmembrane domain (TM2) and that a disulfide bond holds the resulting two fragments together...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100778/structure-of-the-z-ring-associated-protein-zapd-bound-to-the-c-terminal-domain-of-the-tubulin-like-protein-ftsz-suggests-mechanism-of-z-ring-stabilization-through-ftsz-crosslinking
#14
Maria Schumacher, Kuo-Hsiang Huang, Wenjie Zeng, Anuradha Janakiraman
Cell division in most bacteria is mediated by the tubulin-like FtsZ protein, which polymerizes in a GTP-dependent manner to form the cytokinetic Z ring. A diverse repertoire of FtsZ binding proteins affect FtsZ localization and polymerization to ensure correct Z ring formation. Many of these proteins bind the C-terminal domain (CTD) of FtsZ, which serves as a hub for FtsZ regulation. FtsZ ring-associated proteins, ZapA-D (Zaps), are important FtsZ regulatory proteins that stabilize FtsZ assembly and enhance Z ring formation by increasing lateral assembly of FtsZ protofilaments, which then form the Z ring...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100777/structure-activity-relationship-of-%C3%AE-mating-pheromone-from-the-fungal-pathogen-fusarium-oxysporum
#15
Stefania Vitale, Angélica Partida-Hanon, Soraya Serrano, Álvaro Martínez-Del-Pozo, Antonio Di Pietro, David Turrà, Marta Bruix
During sexual development, ascomycete fungi produce two types of peptide pheromones termed a and α. The α pheromone from the budding yeast Saccharomyces cerevisiae, a thirteen residue peptide which elicits cell cycle arrest and chemotropic growth, has served as paradigm for the interaction of small peptides with their cognate G protein-coupled receptors (GPCRs). However, no structural information is currently available for α pheromones from filamentous ascomycetes, which are significantly shorter and share almost no sequence similarity with the S...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100776/crystallographic-snapshots-of-class-a-%C3%AE-lactamase-catalysis-reveal-structural-changes-that-facilitate-%C3%AE-lactam-hydrolysis
#16
Xuehua Pan, Yunjiao He, Jinping Lei, Xuhui Huang, Yanxiang Zhao
β-lactamases confer resistance to β-lactam-based antibiotics. There is great interest in understanding their mechanisms to enable the development of β-lactamase-specific inhibitors. The mechanism of Class A β-lactamases has been studied extensively, revealing Lys73 and Glu166 as general bases that assist the catalytic residue Ser70. However, the specific roles of these two residues within the catalytic cycle remains not fully understood. To help resolve this question, we first identified a Glu166His mutant that is functional but kinetically slowed...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100775/cool-associated-tyrosine-phosphorylated-protein-1-is-required-for-the-anchorage-independent-growth-of-cervical-carcinoma-cells-by-binding-paxillin-and-promoting-akt-activation
#17
Sungsoo M Yoo, Arash Latifkar, Richard A Cerione, Marc A Antonyak
Cool-associated tyrosine phosphorylated protein-1 (Cat-1) is a signaling scaffold, as well as an ADP-ribosylation factor-GTPase-activating protein (ARF-GAP). Although best known for its role in cell migration, we recently showed that the ability of Cat-1 to bind paxillin, a major constituent of focal complexes, was also essential for the anchorage-independent growth of HeLa cervical carcinoma cells. Here, we set out to learn more about the underlying mechanism by which Cat-paxillin interactions mediate this effect...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100774/loss-of-cyclin-dependent-kinase-2-in-the-pancreas-links-primary-%C3%AE-cell-dysfunction-to-progressive-depletion-of-%C3%AE-cell-mass-and-diabetes
#18
So Yoon Kim, Ji-Hyeon Lee, Matthew J Merrins, Oksana Gavrilova, Xavier Bisteau, Philipp Kaldis, Leslie S Satin, Sushil G Rane
The failure of pancreatic islet β-cells is a major contributor to the etiology of Type 2 diabetes. β-cell dysfunction and declining β-cell mass are two mechanisms that contribute to this failure, although it is unclear if they are molecularly linked. Here, we show that the cell cycle regulator, cyclin-dependent kinase 2 (Cdk2), couples primary β-cell dysfunction to the progressive deterioration of β-cell mass in diabetes. Mice with pancreas-specific deletion of Cdk2 are glucose intolerant primarily due to defects in glucose-stimulated insulin secretion...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100773/targeted-fc%C3%AE-r-mediated-clearance-by-a-biparatopic-bispecific-antibody
#19
Srinath Kasturirangan, G Jonah Rainey, Linda Xu, Xinwei Wang, Alyse Portnoff, Tracy Chen, Christine Fazenbaker, Helen Zhong, Jared Bee, Zhutian Zeng, Craig Jenne, Herren Wu, Changshou Gao
Soluble ligands have commonly been targeted by antibody therapeutics for cancers and other diseases. While monoclonal antibodies targeting such ligands can block their interactions with their cognate receptors, they can also significantly increase the half-life of their ligands by FcRn-mediated antibody recycling, thereby evading ligand renal clearance and requiring increasingly high antibody doses to neutralize the increasing pool of target. To overcome this issue, we generated a bispecific/biparatopic antibody (BiSAb) that targets two different epitopes on IL-6 to block IL-6 mediated signaling...
January 18, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28100772/de-novo-sphingolipid-biosynthesis-is-required-for-adipocyte-survival-and-metabolic-homeostasis
#20
Aikaterini Alexaki, Benjamin A Clarke, Oksana Gavrilova, Yinyan Ma, Hongling Zhu, Xinran Ma, Lingyan Xu, Galina Tuymetova, Bridget C Larman, Maria L Allende, Teresa M Dunn, Richard L Proia
Sphingolipids are a diverse class of essential cellular lipids that function as structural membrane components and as signaling molecules. Cells acquire sphingolipids by both de novo biosynthesis and recycling of exogenous sphingolipids. The individual importance of these pathways for the generation of essential sphingolipids in differentiated cells is not well understood. To investigate the requirement for de novo sphingolipid biosynthesis in adipocytes, a cell type with highly regulated lipid metabolism, we generated mice with an adipocyte-specific deletion of Sptlc1 Sptlc1 is an obligate subunit of serine palmitoyltransferase, the enzyme responsible for the first and rate-limiting step of de novo sphingolipid biosynthesis...
January 18, 2017: Journal of Biological Chemistry
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