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https://www.readbyqxmd.com/read/28109288/long-intergenic-non-coding-rna-00152-promotes-lung-adenocarcinoma-proliferation-via-interacting-with-ezh2-and-repressing-il24-expression
#1
Qin-Nan Chen, Xin Chen, Zhen-Yao Chen, Feng-Qi Nie, Chen-Chen Wei, Hong-Wei Ma, Li Wan, Shuai Yan, Sheng-Nan Ren, Zhao-Xia Wang
BACKGROUND: Numerous studies have shown that long non-coding RNAs (lncRNAs) behave as a novel class of transcript during multiple cancer processes, such as cell proliferation, apoptosis, migration, and invasion. LINC00152 is located on chromosome 2p11.2, and has a transcript length of 828 nucleotides. The biological role of LINC00152 in LAD(lung adenocarcinoma) remains unknown. METHODS: Quantitative reverse transcription PCR(qRT-PCR) was used to detect LINC00152 expression in 60 human LAD tissues and paired normal tissues...
January 21, 2017: Molecular Cancer
https://www.readbyqxmd.com/read/28108261/heterotrimeric-g-protein-g%C3%AE-s-subunit-attenuates-plekhg2-a-rho-family-specific-guanine-nucleotide-exchange-factor-by-direct-interaction
#2
Kazue Sugiyama, Kenji Tago, Sayumi Matsushita, Masashi Nishikawa, Katsuya Sato, Yoshinori Muto, Takahiro Nagase, Hiroshi Ueda
PLEKHG2 is a Gβγ-dependent guanine nucleotide exchange factor (GEF) for the small GTPases Rac and Cdc42, and has been shown to mediate signalling pathways such as actin cytoskeleton reorganization and serum response element (SRE)-dependent gene transcription. Here we show that the constitutively active mutant of the Gαs subunit significantly attenuated PLEKHG2-induced SRE-mediated gene transcription. Strikingly, we observed that the constitutive activation of endogenous Gαs by treatment with CTx caused a similar inhibitory effect on PLEKHG2-induced activation of SRE...
January 17, 2017: Cellular Signalling
https://www.readbyqxmd.com/read/28103901/pathogenic-lrrk2-variants-are-gain-of-function-mutations-that-enhance-lrrk2-mediated-repression-of-%C3%AE-catenin-signaling
#3
Daniel C Berwick, Behzad Javaheri, Andrea Wetzel, Mark Hopkinson, Jonathon Nixon-Abell, Simone Grannò, Andrew A Pitsillides, Kirsten Harvey
BACKGROUND: LRRK2 mutations and risk variants increase susceptibility to inherited and idiopathic Parkinson's disease, while recent studies have identified potential protective variants. This, and the fact that LRRK2 mutation carriers develop symptoms and brain pathology almost indistinguishable from idiopathic Parkinson's disease, has led to enormous interest in this protein. LRRK2 has been implicated in a range of cellular events, but key among them is canonical Wnt signalling, which results in increased levels of transcriptionally active β-catenin...
January 19, 2017: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/28096388/competition-of-calcified-calmodulin-n-lobe-and-pip2-to-an-lqt-mutation-site-in-kv7-1-channel
#4
William Sam Tobelaim, Meidan Dvir, Guy Lebel, Meng Cui, Tal Buki, Asher Peretz, Milit Marom, Yoni Haitin, Diomedes E Logothetis, Joel Alan Hirsch, Bernard Attali
Voltage-gated potassium 7.1 (Kv7.1) channel and KCNE1 protein coassembly forms the slow potassium current IKS that repolarizes the cardiac action potential. The physiological importance of the IKS channel is underscored by the existence of mutations in human Kv7.1 and KCNE1 genes, which cause cardiac arrhythmias, such as the long-QT syndrome (LQT) and atrial fibrillation. The proximal Kv7.1 C terminus (CT) binds calmodulin (CaM) and phosphatidylinositol-4,5-bisphosphate (PIP2), but the role of CaM in channel function is still unclear, and its possible interaction with PIP2 is unknown...
January 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28063493/measuring-diacylglycerol-kinase-%C3%AE-activity-and-binding
#5
B Tu-Sekine, D M Raben
This section provides detailed protocols for the analysis of a mammalian diacylglycerol kinase, DGKθ, including an activity assay, a kinetic analysis, preparation of small unilamellar vesicles, and a vesicle pulldown assay. The goal of this section is to provide an overview of the unique challenges inherent in the study of an interfacial enzyme such as DGKθ and to outline methods useful for analysis. We include a short tutorial on selecting lipids for forming the interface since this is critical for a successful in vitro assay, and lipids are important regulators of this enzyme...
2017: Methods in Enzymology
https://www.readbyqxmd.com/read/28041939/involvement-of-the-coatomer-protein-complex-i-in-the-intracellular-traffic-of-the-delta-opioid-receptor
#6
Étienne St-Louis, Jade Degrandmaison, Sébastien Grastilleur, Samuel Génier, Véronique Blais, Christine Lavoie, Jean-Luc Parent, Louis Gendron
The delta opioid receptor (DOPr) is known to be mainly expressed in intracellular compartments. It remains unknown why DOPr is barely exported to the cell surface, but it seems that a substantial proportion of the immature receptor is trapped within the endoplasmic reticulum (ER) and the Golgi network. In the present study, we performed LC-MS/MS analysis to identify putative protein partners involved in the retention of DOPr. Analysis of the proteins co-immunoprecipitating with Flag-DOPr in transfected HEK293 cells revealed the presence of numerous subunits of the coatomer protein complex I (COPI), a vesicle-coating complex involved in recycling resident proteins from the Golgi back to the ER...
December 30, 2016: Molecular and Cellular Neurosciences
https://www.readbyqxmd.com/read/28028182/domain-mapping-of-the-heat-shock-protein-70-reveals-that-glutamic-acid-446-and-arginine-447-are-critical-for-regulating-superoxide-dismutase-2-function
#7
Adeleye J Afolayan, Maxwell Alexander, Rebecca L Holme, Teresa Michalkiewicz, Ujala Rana, Ru-Jeng Teng, Sara Zemanovic, Daisy Sahoo, Kirkwood A Pritchard, Girija G Konduri
Stress-inducible heat shock protein 70 (hsp70) interacts with superoxide dismutase-2 (SOD2) in the cytosol after synthesis to transfer the enzyme to the mitochondria for subsequent activation. However, the structural basis for this interaction remains to be defined. To map the SOD2-binding site in hsp70, mutants of hsp70 were made and tested for their ability to bind SOD2. These studies showed that SOD2 binds in the amino acid 393-537 region of the chaperone. To map the hsp70-binding site in SOD2, we used a series of pulldown assays and showed that hsp70 binds to the amino-terminal domain of SOD2...
December 27, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28018022/human-ribosomal-proteins-rpel27-rpel43-and-rpel41-are-upregulated-in-nasopharyngeal-carcinoma-cell-lines
#8
Edmund Ui-Hang Sim, Stella Li-Li Chan, Kher-Lee Ng, Choon-Weng Lee, Kumaran Narayanan
Apart from their canonical role in ribosome biogenesis, there is increasing evidence of ribosomal protein genes' involvement in various cancers. A previous study by us revealed significant differential expression of three ribosomal protein genes (RPeL27, RPeL41, and RPeL43) between cell lines derived from tumor and normal nasopharyngeal epithelium. However, the results therein were based on a semiquantitative assay, thus preliminary in nature. Herein, we provide findings of a deeper analysis of these three genes in the context to nasopharyngeal carcinoma (NPC) tumorigenesis...
2016: Disease Markers
https://www.readbyqxmd.com/read/28008132/conserved-residues-in-ycf54-are-required-for-protochlorophyllide-formation-in-synechocystis-sp-pcc-6803
#9
Sarah Hollingshead, Sophie Bliss, Patrick J Baker, C Neil Hunter
Chlorophylls are modified tetrapyrrole molecules, essential for photosynthesis. These pigments possess an isocyclic E ring formed by the Mg-protoporphyrin IX monomethylester cyclase (MgPME-cyclase). We assessed the in vivo effects of altering seven highly conserved residues within Ycf54, which is required for MgPME-cyclase activity in the cyanobacterium Synechocystis Synechocystis strains harbouring the Ycf54 alterations D39A, F40A and R82A were blocked to varying degrees at the MgPME-cyclase step, whereas the A9G mutation reduced Ycf54 levels by ~75%...
December 22, 2016: Biochemical Journal
https://www.readbyqxmd.com/read/28002732/methyl-cpg-mbd2-interaction-requires-minimum-separation-and-exhibits-minimal-sequence-specificity
#10
Blythe Moreland, Kenji Oman, John Curfman, Pearlly Yan, Ralf Bundschuh
Determining the pattern of methylation at CpG dinucleotides in a cell remains an essential component of epigenetic profiling. The correlations among methylation, gene expression, and accompanying disease have just begun to be explored. Many experiments for sensing methylation use a relatively inexpensive, high-throughput approach with a methyl-binding domain (MBD) protein that preferentially binds to methylated CpGs. Here, we characterize the cooperativity and sequence specificity of MBD2-DNA binding in a pulldown experiment revealing three potential biases in such experiments...
December 20, 2016: Biophysical Journal
https://www.readbyqxmd.com/read/27997899/bufalin-inhibits-htert-expression-and-colorectal-cancer-cell-growth-by-targeting-cpsf4
#11
Ningning Zhang, Yunpeng Xie, Yidi Tai, Yingying Gao, Wei Guo, Wendan Yu, Jia Li, Xu Feng, Jiaojiao Hao, Yue Gao, Xinrui Zhao, Yina Liao, Wei Jiang, Ge Liu, Wuguo Deng, Xiaonan Cui
BACKGROUND/AIMS: Bufalin can induce apoptosis in certain human cancer cell lines, but bufalin has not yet been thoroughly evaluated in colorectal cancer cells. Cleavage and polyadenylation specific factor 4 (CPSF4) and human telomerase reverse transcriptase (hTERT) play important roles in colorectal cancer growth. The aim of this study was to investigate the roles and interactions of bufalin, CPSF4 and hTERT and the effects of bufalin in human colorectal cancer. METHODS: We treated LoVo and SW620 cells with bufalin to investigate the effect of bufalin on proliferation, apoptosis and migration...
2016: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/27991487/the-non-canonical-mitochondrial-inner-membrane-presequence-translocase-of-trypanosomatids-contains-two-essential-rhomboid-like-proteins
#12
Anke Harsman, Silke Oeljeklaus, Christoph Wenger, Jonathan L Huot, Bettina Warscheid, André Schneider
Mitochondrial protein import is essential for all eukaryotes. Here we show that the early diverging eukaryote Trypanosoma brucei has a non-canonical inner membrane (IM) protein translocation machinery. Besides TbTim17, the single member of the Tim17/22/23 family in trypanosomes, the presequence translocase contains nine subunits that co-purify in reciprocal immunoprecipitations and with a presequence-containing substrate that is trapped in the translocation channel. Two of the newly discovered subunits are rhomboid-like proteins, which are essential for growth and mitochondrial protein import...
December 19, 2016: Nature Communications
https://www.readbyqxmd.com/read/27984101/imaging-flow-cytometry-and-gst-pulldown-assays-provide-new-insights-into-channel-catfish-leukocyte-immune-type-receptor-mediated-phagocytic-pathways
#13
Myron A Zwozdesky, Chenjie Fei, Dustin M E Lillico, James L Stafford
Channel catfish (Ictalurus punctatus) leukocyte immune-type receptors (IpLITRs) control various innate immune cell effector responses including the phagocytic process. This large immunoregulatory receptor family also consists of multiple receptor-types with variable signaling abilities that is dependent on their inherent or acquired tyrosine-containing cytoplasmic tail (CYT) regions. For example, IpLITR 2.6b associates with the immunoreceptor tyrosine-based activation motif (ITAM)-containing adaptor molecule IpFcRγ-L, and when expressed in mammalian cells it activates phagocytosis using a similar profile of intracellular signaling mediators that also regulate the prototypical mammalian Fc receptor (FcR) phagocytic pathway...
October 27, 2016: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/27934296/identification-of-candidate-host-cell-factors-required-for-actin-based-motility-of-burkholderia-pseudomallei
#14
Niramol Jitprasutwit, Nurhamimah Zainal-Abidin, Charles Vander Broek, Dominic Kurian, Sunee Korbsrisate, Mark P Stevens, Joanne M Stevens
Intracellular actin-based motility of the melioidosis pathogen Burkholderia pseudomallei requires the bacterial factor BimA. Located at one pole of the bacterium, BimA recruits and polymerizes cellular actin to promote bacterial motility within and between cells. Here, we describe an affinity approach coupled with mass spectrometry to identify cellular proteins recruited to BimA-expressing bacteria under conditions that promote actin polymerization. We identified a group of cellular proteins that are recruited to the B...
December 2, 2016: Journal of Proteome Research
https://www.readbyqxmd.com/read/27932498/harnessing-molecular-motors-for-nanoscale-pulldown-in-live-cells
#15
Jonathan E Bird, Melanie Barzik, Meghan C Drummond, Daniel C Sutton, Spencer M Goodman, Eva L Morozko, Stacey M Cole, Alexandra K Boukhvalova, Jennifer Skidmore, Diana Syam, Elizabeth A Wilson, Tracy Fitzgerald, Atteeq U Rehman, Donna M Martin, Erich T Boger, Inna A Belyantseva, Thomas B Friedman
Protein-protein interactions (PPIs) regulate assembly of macromolecular complexes, yet these remain challenging to study within the native cytoplasm where they normally exert their biological effect. Here, we miniaturize the concept of affinity pulldown, a gold-standard in vitro PPI interrogation technique, to perform nanoscale pulldowns (NanoSPD) within living cells. NanoSPD hijacks the normal process of intracellular trafficking by myosin motors to forcibly pull fluorescently-tagged protein complexes along filopodial actin filaments...
December 8, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27928058/identification-of-senescence-associated-circular-rnas-sac-rnas-reveals-senescence-suppressor-circpvt1
#16
Amaresh C Panda, Ioannis Grammatikakis, Kyoung Mi Kim, Supriyo De, Jennifer L Martindale, Rachel Munk, Xiaoling Yang, Kotb Abdelmohsen, Myriam Gorospe
Using RNA sequencing (RNA-Seq), we compared the expression patterns of circular RNAs in proliferating (early-passage) and senescent (late-passage) human diploid WI-38 fibroblasts. Among the differentially expressed senescence-associated circRNAs (which we termed 'SAC-RNAs'), we identified CircPVT1, generated by circularization of an exon of the PVT1 gene, as a circular RNA showing markedly reduced levels in senescent fibroblasts. Reducing CircPVT1 levels in proliferating fibroblasts triggered senescence, as determined by a rise in senescence-associated β-galactosidase activity, higher abundance of CDKN1A/P21 and TP53, and reduced cell proliferation...
December 7, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27914499/exercise-training-improves-exercise-capacity-and-quality-of-life-after-transcatheter-aortic-valve-implantation-a-randomized-pilot-trial
#17
Axel Pressler, Jeffrey W Christle, Benjamin Lechner, Viola Grabs, Bernhard Haller, Ina Hettich, David Jochheim, Julinda Mehilli, Rüdiger Lange, Sabine Bleiziffer, Martin Halle
BACKGROUND: Transcatheter aortic valve implantation (TAVI) is increasingly applied for aortic stenosis in elderly patients with impaired mobility and reduced quality of life. These patients may particularly benefit from postinterventional exercise programs, but no randomized study has evaluated the safety and efficacy of exercise in this population. METHODS: In a prospective pilot study, 30 patients after TAVI (mean age, 81±6 years, 44% female, 83±34 days postintervention) were randomly allocated 1:1 to a training group (TG) performing 8 weeks of supervised combined endurance and resistance exercise or to usual care...
December 2016: American Heart Journal
https://www.readbyqxmd.com/read/27893478/loads-and-movement-speeds-dictate-differences-in-power-output-during-circuit-training
#18
Kirk B Roberson, Sean Chowdhari, Morgan White, Joseph F Signorile
Power training has become a common exercise intervention for improving muscle strength, power, and physical function, while reducing injury risk. Few studies, however, have evaluated acute load changes on power output during traditional resistance-training protocols. Therefore, the aim of this study was to quantify the effects of different loading patterns on power output during a single session of circuit resistance training (CRT). Nine males (age=19.4±0.9 y) and eleven females (age=20.6±1.6 y) completed three CRT protocols during separate testing sessions using seven pneumatic exercises...
November 16, 2016: Journal of Strength and Conditioning Research
https://www.readbyqxmd.com/read/27887846/long-non-coding-rna-metastasis-associated-lung-adenocarcinoma-transcript-1-regulates-the-expression-of-gli2-by-mir-202-to-strengthen-gastric-cancer-progression
#19
Yue Zhang, Zhuo Chen, Ming-Jing Li, Huan-Yu Guo, Nian-Cai Jing
BACKGROUND: Gastric cancer (GC) is one of the most common malignancies and ranks the second leading cause of cancer death worldwide. Some studies had reported the tumor-promoting effects of long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) as a competing endogenous RNA (ceRNA) by sponging to microRNAs. However, the molecular mechanism of ceRNA regulatory pathway involving MALAT1 in GC remains unclear. METHODS: MALAT1 and miR -202 expression was detected by quantitative real time PCR (qRT-PCR) in 60 gastric cancer tissues and adjacent normal tissues, CCK8 cell proliferation assays, cell cycle analysis and cell apoptosis assays were performed to detect the GC cell proliferation and apoptosis...
January 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/27881679/the-shigella-virulence-factor-icsa-relieves-n-wasp-autoinhibition-by-displacing-the-verprolin-homology-cofilin-acidic-vca-domain
#20
Rui P M Mauricio, Cy M Jeffries, Dmitri I Svergun, Janet E Deane
Shigella flexneri is a bacterial pathogen that invades cells of the gastrointestinal tract, causing severe dysentery. Shigella mediates intracellular motility and spreading via actin comet tail formation. This process is dependent on the surface-exposed, membrane-embedded virulence factor IcsA, which recruits the host actin regulator N-WASP. Although it is clear that Shigella requires N-WASP for this process, the molecular details of this interaction and the mechanism of N-WASP activation remain poorly understood...
January 6, 2017: Journal of Biological Chemistry
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