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https://www.readbyqxmd.com/read/27934698/ltr-mediated-retroposition-as-a-mechanism-of-rna-based-duplication-in-metazoans
#1
Shengjun Tan, Margarida Cardoso-Moreira, Wenwen Shi, Dan Zhang, Jiawei Huang, Yanan Mao, Hangxing Jia, Yaqiong Zhang, Chunyan Chen, Yi Shao, Liang Leng, Zhonghua Liu, Xun Huang, Manyuan Long, Yong E Zhang
In a broad range of taxa, genes can duplicate through an RNA intermediate in a process mediated by retrotransposons (retroposition). In mammals, L1 retrotransposons drive retroposition, but the elements responsible for retroposition in other animals have yet to be identified. Here, we examined young retrocopies from various animals that still retain the sequence features indicative of the underlying retroposition mechanism. In Drosophila melanogaster, we identified and de novo assembled 15 polymorphic retrocopies and found that all retroposed loci are chimeras of internal retrocopies flanked by discontinuous LTR retrotransposons...
December 2016: Genome Research
https://www.readbyqxmd.com/read/27934684/performing-yeast-one-hybrid-library-screens
#2
Juan I Fuxman Bass, John S Reece-Hoyes, Albertha J M Walhout
Yeast one-hybrid (Y1H) assays are used to identify which transcription factor (TF) "prey" molecules can bind a DNA fragment of interest that is used as "bait". Y1H assays involve introducing plasmids that encode TFs into a yeast "bait strain" in which the DNA fragment of interest is integrated upstream of one or more reporters, and activation of these reporters indicates that a TF-DNA interaction has occurred. These plasmids express each TF as a hybrid protein (hence the "one-hybrid" name) fused to the activation domain (AD) of the yeast TF Gal4...
December 1, 2016: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/27934109/in-vivo-biotransformation-of-the-fusion-protein-tetranectin-apolipoprotein-a1-analyzed-by-ligand-binding-mass-spectrometry-combined-with-quantitation-by-elisa
#3
Manfred Zell, Christophe Husser, Roland F Staack, Gregor Jordan, Wolfgang F Richter, Simone Schadt, Axel Pähler
The in vivo biotransformation of a novel fusion protein tetranectin/apolipoprotein A1 (TN-ApoA1) was investigated by ligand-binding mass spectrometry (LB-MS) in support of enzyme-linked immunosorbent assays (ELISA). The main focus was on catabolites formed by proteolysis of the fusion protein in rabbit following intravenous administration of lipidated TN-ApoA1. The drug and its catabolites were isolated from rabbit plasma by immunocapture with a monoclonal antibody (mAb) binding to the fusion region of TN-ApoA1...
December 6, 2016: Analytical Chemistry
https://www.readbyqxmd.com/read/27933781/structure-and-function-of-the-hepatitis-c-virus-envelope-glycoproteins-e1-and-e2-antiviral-and-vaccine-targets
#4
Holly Freedman, Michael R Logan, John Lok Man Law, Michael Houghton
The hepatitis C virus (HCV) envelope glycoproteins E1 and E2 are critical in viral attachment and cell fusion, and studies of these proteins may provide valuable insights into their potential uses in vaccines and antiviral strategies. Progress has included elucidating the crystal structures of portions of their ectodomains, as well as many other studies of hypervariable regions, stem regions, glycosylation sites, and the participation of E1/E2 in viral fusion with the endosomal membrane. The available structural data have shed light on the binding sites of cross-neutralizing antibodies...
November 11, 2016: ACS Infectious Diseases
https://www.readbyqxmd.com/read/27933482/effect-of-linker-length-and-flexibility-on-the-clostridium-thermocellum-esterase-displayed-on-bacillus-subtilis-spores
#5
Huayou Chen, Bangguo Wu, Tianxi Zhang, Jinru Jia, Jian Lu, Zhi Chen, Zhong Ni, Tianwei Tan
In fusion protein design strategies, the flexibility and length of linkers are important parameters affecting the bioactivity of multifunctional proteins. A series of fusion proteins with different linkers were constructed. The effect of temperature, pH, and organic solvents was investigated on the enzymatic activity. Fusion proteins with P1(PTPTPT) and P2((PTPTPT)2) linkers remained highly active with wide temperature range. At pH 9.6, the relative activity of fusion proteins with (PTPTPT)2 and S2(EGKSSGSGSESKST) linkers was 70 and 62 % (1...
December 8, 2016: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/27933304/preclinical-validation-lv-il-12-transduction-of-patient-leukemia-cells-for-immunotherapy-of-aml
#6
Ju Huang, Yuanfeng Liu, Bryan C Au, Dwayne L Barber, Andrea Arruda, Axel Schambach, Michael Rothe, Mark D Minden, Christopher J Paige, Jeffrey A Medin
Interleukin-12 (IL-12) is a potent cytokine that may be harnessed to treat cancer. To date, nearly 100 IL-12-based clinical trials have been initiated worldwide. Yet systemic administration of IL-12 is toxic. Different strategies are being developed to reduce such toxicities by restricting IL-12 distribution. Our previous studies employed lentivector-mediated expression of murine IL-12 in tumor cells and demonstrated effective protection in both mouse leukemia and solid tumor challenge models. In this study, we carried out preclinical validation studies using a novel lentivector to engineer expression of human IL-12 in acute myeloid leukemia blast cells isolated from 21 patients...
2016: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/27933076/overexpression-of-tanac2d-displays-opposite-responses-to-abiotic-stresses-between-seedling-and-mature-stage-of-transgenic-arabidopsis
#7
Quanjun Huang, Yan Wang
Environmental stresses frequently affect plant growth and development, and many genes have been found to be induced by unfavorable environmental conditions. Here, we reported the biological functions of TaNAC2D, a stress-related NAC (NAM, ATAF, and CUC) gene from wheat. TaNAC2D showed transcriptional activator activity in yeast. TaNAC2D-GFP fusion protein was localized in the nucleus of wheat mesophyll protoplasts. TaNAC2D transcript abundance was significantly induced by NaCl, PEG6000, and abscisic acid (ABA) at seedling stage, and repressed by NaCl and PEG6000 at mature plant stage...
2016: Frontiers in Plant Science
https://www.readbyqxmd.com/read/27932952/evaluating-tools-for-live-imaging-of-structural-plasticity-at-the-axon-initial-segment
#8
Adna S Dumitrescu, Mark D Evans, Matthew S Grubb
The axon initial segment (AIS) is a specialized neuronal compartment involved in the maintenance of axo-dendritic polarity and in the generation of action potentials. It is also a site of significant structural plasticity-manipulations of neuronal activity in vitro and in vivo can produce changes in AIS position and/or size that are associated with alterations in intrinsic excitability. However, to date all activity-dependent AIS changes have been observed in experiments carried out on fixed samples, offering only a snapshot, population-wide view of this form of plasticity...
2016: Frontiers in Cellular Neuroscience
https://www.readbyqxmd.com/read/27932492/novel-regulatory-roles-of-mff-and-drp1-in-e3-ubiquitin-ligase-march5-dependent-degradation-of-mid49-and-mcl1-and-control-of-mitochondrial-dynamics
#9
Edward Cherok, Shan Xu, Sunan Li, Shweta Das, W Alex Meltzer, Michal Zalzman, Chunxin Wang, Mariusz Karbowski
MARCH5, an OMM-associated E3 ubiquitin ligase, controls mitochondrial function. Despite its importance, the mechanism and factors controlling MARCH5 activity are largely unknown. Here we report that the MARCH5 C-terminal domain plays a critical role in degradation of MARCH5 substrates, likely by facilitating release of ubiquitinated proteins from the OMM. We also found that the mitochondrial fission proteins Drp1 and Mff negatively regulate MARCH5's activity toward MiD49 and Mcl1. Knockouts of either Drp1 or Mff led to reduced expression, shorter half-lives, and increased ubiquitination of MiD49 and Mcl1...
December 8, 2016: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27932448/dedicated-snares-and-specialized-trim-cargo-receptors-mediate-secretory-autophagy
#10
Tomonori Kimura, Jingyue Jia, Suresh Kumar, Seong Won Choi, Yuexi Gu, Michal Mudd, Nicolas Dupont, Shanya Jiang, Ryan Peters, Farzin Farzam, Ashish Jain, Keith A Lidke, Christopher M Adams, Terje Johansen, Vojo Deretic
Autophagy is a process delivering cytoplasmic components to lysosomes for degradation. Autophagy may, however, play a role in unconventional secretion of leaderless cytosolic proteins. How secretory autophagy diverges from degradative autophagy remains unclear. Here we show that in response to lysosomal damage, the prototypical cytosolic secretory autophagy cargo IL-1β is recognized by specialized secretory autophagy cargo receptor TRIM16 and that this receptor interacts with the R-SNARE Sec22b to recruit cargo to the LC3-II(+) sequestration membranes...
December 8, 2016: EMBO Journal
https://www.readbyqxmd.com/read/27932421/myb89-transcription-factor-represses-seed-oil-accumulation
#11
Dong Li, Changyu Jin, Shaowei Duan, Yana Zhu, Shuanghui Qi, Kaige Liu, Chenhao Gao, Haoli Ma, Meng Zhang, Yuncheng Liao, Mingxun Chen
In many higher plants, seed oil accumulation is precisely controlled by intricate multi-level regulatory networks, among which transcriptional regulation mainly influences oil biosynthesis. In Arabidopsis, the master positive transcription factors, WRINKLED1 (WRI1) and LEAFY COTYLEDON1-LIKE (L1L), are important for seed oil accumulation. We found that an R2R3-MYB transcription factor, MYB89, was predominantly expressed in developing seeds during maturation. Oil and major fatty acid biosynthesis in seeds was significantly promoted by myb89-1 mutation and MYB89 knockdown; MYB89 was thus an important repressor during seed oil accumulation...
December 8, 2016: Plant Physiology
https://www.readbyqxmd.com/read/27930268/recombinant-bacillus-subtilis-spores-expressing-cholera-toxin-b-subunit-and-helicobacter-pylori-urease-b-confers-protection-against-h-pylori-in-mice
#12
Zhenwen Zhou, Hui Dong, Shuwen Yao, Yanmei Huang, Bingshao Liang, Yongqiang Xie, Yan Long, Jialiang Mai, Sitang Gong
Helicobacter pylori infection is associated with chronic gastritis, peptic ulcers, gastric cancer and mucosa-associated lymphoid tissue lymphoma. The limitations of current therapies for Helicobacter pylori infection include poor compliance and antibiotic resistance. Therefore, an effective anti-H. pylori vaccine would be an alternative or complement to antibiotic treatment. Urease B (UreB) is considered an ideal vaccine antigen against H. pylori infection. In this study, cholera toxin B subunit (CTB), a mucosal adjuvant, was used to enhance the immunogenicity of novel Bacillus subtilis spores vaccine that express CTB-UreB along with the B...
December 8, 2016: Journal of Medical Microbiology
https://www.readbyqxmd.com/read/27929754/hpv-16-e5-oncoprotein-is-expressed-in-early-stage-carcinogenesis-and-can-be-a-target-of-immunotherapy
#13
Paolini Francesca, Curzio Gianfranca, Marcelo Nazario Cordeiro, Silvia Massa, Luciano Mariani, Fulvia Pimpinelli, Antonio Carlos de Freitas, Rosella Franconi, Aldo Venuti
HPV16 persistent infection is a well-known condition that precedes human cancer development. High risk HPV E5 proteins cooperate with E6/E7 oncogenes to promote hyper-proliferation of infected cells leading to possible cancer progression. Thus, presence of E5 viral transcripts could be a key marker of active infection and, in turn, a target of immunotherapy. Purpose of the study is to detect E5 transcripts in clinical samples and to explore the activity of novel anti-HPV16 E5 DNA vaccines. HPV transcripts were detected by PCR with specific primers encompassing the splice-donor sites of E5 transcript...
December 8, 2016: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/27929705/snapin-is-critical-for-lysosomal-acidification-and-autophagosome-maturation-in-macrophages
#14
Bo Shi, Qi-Quan Huang, Robert Birkett, Renee Doyle, Andrea Dorfleutner, Christian Stehlik, Congcong He, Richard M Pope
We previously observed that SNAPIN, which is an adaptor protein in the SNARE core complex, was highly expressed in rheumatoid arthritis synovial tissue macrophages, but its role in macrophages and autoimmunity is unknown. To identify SNAPIN's role in these cells, we employed siRNA to silence the expression of SNAPIN in primary human macrophages. Silencing SNAPIN resulted in swollen lysosomes with impaired CTSD (cathepsin D) activation, although total CTSD was not reduced. Neither endosome cargo delivery nor lysosomal fusion with endosomes or autophagosomes was inhibited following the forced silencing of SNAPIN...
December 8, 2016: Autophagy
https://www.readbyqxmd.com/read/27928921/discovery-of-natural-mouse-serum-derived-hiv-1-entry-inhibitor-s
#15
M Wei, Y Chen, J Xi, S Ru, M Ji, D Zhang, Q Fang, B Tang
Among rationally designed human immunodeficiency virus 1 (HIV-1) inhibitors, diverse natural factors have showed as potent anti-HIV activity in human blood. We have discovered that the boiled supernatant of healthy mouse serum could suppress HIV-1 entry, and exhibited reduced inhibitory activity after trypsin digestion. Further analysis demonstrated that only the fraction containing 10-25 K proteins could inhibit HIV-1 mediated cell-cell fusion. These results suggest that the 10-25 K protein(s) is novel natural HIV-1 entry inhibitor(s)...
2016: Acta Virologica
https://www.readbyqxmd.com/read/27928159/effect-of-trem-1-blockade-and-single-nucleotide-variants-in-experimental-renal-injury-and-kidney-transplantation
#16
A Tammaro, J Kers, D Emal, I Stroo, G J D Teske, L M Butter, N Claessen, J Damman, M Derive, G Navis, S Florquin, J C Leemans, M C Dessing
Renal ischemia reperfusion (IR)-injury induces activation of innate immune response which sustains renal injury and contributes to the development of delayed graft function (DGF). Triggering receptor expressed on myeloid cells-1 (TREM-1) is a pro-inflammatory evolutionary conserved pattern recognition receptor expressed on a variety of innate immune cells. TREM-1 expression increases following acute and chronic renal injury. However, the function of TREM-1 in renal IR is still unclear. Here, we investigated expression and function of TREM-1 in a murine model of renal IR using different TREM-1 inhibitors: LP17, LR12 and TREM-1 fusion protein...
December 8, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27927993/suppression-of-extracellular-invertase-inhibitor-gene-expression-improves-seed-weight-in-soybean-glycine-max
#17
Xiaofei Tang, Tao Su, Mei Han, Lai Wei, Weiwei Wang, Zhiyuan Yu, Yongguo Xue, Hongbin Wei, Yejie Du, Steffen Greiner, Thomas Rausch, Lijun Liu
Cell wall invertase (CWI) and vacuolar invertase (VI) play multiple functions in plant growth. As well as depending on transcriptional and post-transcriptional regulation, there is growing evidence that CWI and VI are also subject to post-translational control by small inhibitory proteins. Despite the significance of this, genes encoding inhibitors, their molecular and biochemical properties, and their potential roles in regulating seed production have not been well documented in soybean (Glycine max). In this study, two invertase inhibitor isoforms, GmCIF1 and GmC/VIF2, were characterized to possess inhibitory activities in vitro via heterologous expression...
December 7, 2016: Journal of Experimental Botany
https://www.readbyqxmd.com/read/27927752/vesicular-ptdins-3-4-5-p3-and-rab-7-are-key-effectors-of-zygote-nuclear-membrane-fusion
#18
Marta G Lete, Richard D Byrne, Alicia Alonso, Dominic Poccia, Banafshé Larijani
Regulation of nuclear envelope (NE) dynamics is an important example of the universal phenomena of membrane fusion. The signalling molecules involved in nuclear membrane fusion may also be conserved in the formation of both pronuclear and zygote NEs in the fertilised egg. Here, we determine that Class I PI3-kinases are needed for in vitro nuclear envelope formation. We show that, in vivo, PtdIns(3,4,5)P3 is transiently located in vesicles around the male pronucleus at the time of nuclear envelope formation and around male and female pronuclei prior to membrane fusion...
December 7, 2016: Journal of Cell Science
https://www.readbyqxmd.com/read/27927751/rab2a-and-rab27a-cooperatively-regulate-transition-from-granule-maturation-to-exocytosis-through-the-dual-effector-noc2
#19
Kohichi Matsunaga, Masato Taoka, Toshiaki Isobe, Tetsuro Izumi
Exocytosis of secretory granules entails budding from the trans-Golgi network, sorting and maturation of cargo proteins, and trafficking and fusion to the plasma membrane. Rab27a regulates the late steps in this process, such as granule recruitment to the fusion site, whereas Rab2a functions in the early steps, such as granule biogenesis and maturation. Here, we demonstrate that these two small GTPases simultaneously bind to Noc2 in GTP-dependent manners, although Rab2a binds only after Rab27a has bound. In pancreatic beta cells, the ternary Rab2a-Noc2-Rab27a complex specifically localizes on perinuclear immature granules, whereas the binary Noc2-Rab27a complex localizes on peripheral mature granules...
December 7, 2016: Journal of Cell Science
https://www.readbyqxmd.com/read/27927521/reoperation-rates-after-long-posterior-spinal-fusion-use-of-recombinant-bone-morphogenetic-protein-in-idiopathic-and-non-idiopathic-scoliosis
#20
Justin C Paul, Baron S Lonner, Shaleen Vira, Ian David Kaye, Thomas J Errico
INTRODUCTION: Bone morphogenetic protein (BMP) can increase the likelihood of solid arthrodesis in spinal surgery. This would imply fewer reoperations for pseudarthrosis, but small cohort sizes are inadequate to monitor these events. We sought to examine adolescent idiopathic (AIS) and non-idiopathic scoliosis (NIS) for reoperation events with and without the use of BMP using a large statewide database. METHODS: The 2008-2011 New York State Inpatient Database was queried using International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes...
July 2016: Spine Deformity
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