FEBS Journal

Cover image for Vol. 281 Issue 7

April 2014

Volume 281, Issue 7

Pages i–iii, 1717–1930

  1. Front Cover

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. You have free access to this content
      Front Cover (page i)

      Article first published online: 1 APR 2014 | DOI: 10.1111/febs.12645

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      Localization of the gaps between the active site of heavily mutated patient-derived HIV-1 proteases and anti-HIV drug darunavir by Milan Kožíšek et al. (pp. 1834–1847).

  2. Editorial Information

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. You have free access to this content
      Editorial Information (pages ii–iii)

      Article first published online: 1 APR 2014 | DOI: 10.1111/febs.12645_1

  3. Editor's Choice

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. You have free access to this content
      Dual sgRNAs facilitate CRISPR/Cas9-mediated mouse genome targeting (pages 1717–1725)

      Jiankui Zhou, Jianying Wang, Bin Shen, Li Chen, Yang Su, Jing Yang, Wensheng Zhang, Xuemei Tian and Xingxu Huang

      Article first published online: 26 FEB 2014 | DOI: 10.1111/febs.12735

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      Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) endonuclease, also known as an RNA-guided endonuclease (RGEN), is increasingly being used for eukaryote genome targeting; however, its efficiency, specificity and versatility needs to be optimized. In this issue, Zhou et al. report an improved method for CRISPR/Cas9-mediated gene targeting, and show that the simultaneous use of dual sgRNAs to target an individual mouse gene is a more efficient and specific method than using just a single sgRNA. This new dual sgRNAs strategy will facilitate CRISPR/Cas9-mediated mammalian genome targeting.

  4. Original Articles

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. You have full text access to this OnlineOpen article
      N-methylmesoporphyrin IX fluorescence as a reporter of strand orientation in guanine quadruplexes (pages 1726–1737)

      Navin C. Sabharwal, Victoria Savikhin, Joshua R. Turek-Herman, John M. Nicoludis, Veronika A. Szalai and Liliya A. Yatsunyk

      Article first published online: 26 FEB 2014 | DOI: 10.1111/febs.12734

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      N-methylmesoporphyrin IX (NMM) shows promise as a ‘turn-on’ fluorescent probe for detecting and differentiating quadruplex structures on the basis of strand orientation. NMM fluorescence intensity is unaffected by duplex, single-stranded and i-motif DNA. In striking contrast, fluorescence intensity increases of 60-, 40-, and 10-fold were measured for NMM in the presence of parallel-stranded, hybrid, or antiparallel quadruplexes, respectively.

    2. The effects of N-ethyl-N′-methyl imidazolium chloride on the solubility, stability and aggregation of tc-rPA (pages 1738–1749)

      Alexander Tischer, Heiko Pultke, Andrea Topf, Matthew Auton, Christian Lange and Hauke Lilie

      Article first published online: 19 FEB 2014 | DOI: 10.1111/febs.12736

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      The ionic liquid EMIMCl is efficient in promoting the refolding of rPA. Molar concentrations of EMIMCl increase the solubility of native and unfolded proteins due to favorable interactions with amino acid side chains, while the interaction with the peptide backbone is unfavorable. Although a refolding enhancer on one side, high concentrations of EMIMCl result in denaturation and aggregation of proteins.

    3. Long non-coding RNA UCA1 increases chemoresistance of bladder cancer cells by regulating Wnt signaling (pages 1750–1758)

      Yu Fan, Bing Shen, Mingyue Tan, Xinyu Mu, Yan Qin, Fang Zhang and Yong Liu

      Article first published online: 20 FEB 2014 | DOI: 10.1111/febs.12737

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      Cisplatin treatment results in the UCA1 expression in vitro and in vivo. UCA1 overexpression significantly increases the cell viability, whereas UCA1 knockdown reduces the cell viability during cisplatin treatment. UCA1 positively regulates Wnt6 expression in vitro and in vivo. Finally, we demonstrated that UCA1 increases the cisplatin resistance of bladder cancer cell by enhancing the expression of Wnt6.

    4. Crystal structures of two tetrameric β-carbonic anhydrases from the filamentous ascomycete Sordaria macrospora (pages 1759–1772)

      Ronny Lehneck, Piotr Neumann, Daniela Vullo, Skander Elleuche, Claudiu T. Supuran, Ralf Ficner and Stefanie Pöggeler

      Article first published online: 19 FEB 2014 | DOI: 10.1111/febs.12738

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      In this manuscript we determined the crystal structures and the in vitro CO2 hydration activities of the plant-type β-carbonic anhydrases CAS1 and CAS2 of the fungus Sordaria macrospora. The oligomeric state of both proteins is tetrameric. Both CAs are weakly only inhibited by anions making them good candidates for industrial applications.

    5. Acyl-coenzyme A:cholesterol acyltransferase 1 – significance of single-nucleotide polymorphism at residue 526 and the role of Pro347 near the fifth transmembrane domain (pages 1773–1783)

      Li-Hao Huang, Koji Nishi, Song Li, Thomas Ho, Ruhong Dong, Catherine C. Y. Chang and Ta-Yuan Chang

      Article first published online: 24 FEB 2014 | DOI: 10.1111/febs.12739

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      Acyl-coenzyme A:cholesterol acyltransferase 1 (ACAT1) contains 550 residues and is a drug target for atherosclerosis and Alzheimer's disease. At residue 526, an A/G single nucleotide polymorphism exists, resulting in glutamine (Q) or arginine (R). We show that enzymatically ACAT1 Q526 is less active than ACAT1 R526 by 40%. We also show that P347 located near TMD #5 modulates enzyme catalysis.

    6. Mutations to a glycine loop in the catalytic site of human Lon changes its protease, peptidase and ATPase activities (pages 1784–1797)

      Ľuboš Ambro, Vladimír Pevala, Gabriela Ondrovičová, Jana Bellová, Nina Kunová, Eva Kutejová and Jacob Bauer

      Article first published online: 1 MAR 2014 | DOI: 10.1111/febs.12740

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      The proteolytic active site of the ATP-dependent, human mitochondrial Lon protease contains two conserved glycine residues, Gly893 and Gly894, which appear both to be involved in the conformational changes that occur upon substrate binding and to influence ATPase activity. The loop containing these residues must move during digestion of larger protein substrates, but not during cleavage of smaller peptides.

    7. Stochastic galactokinase expression underlies GAL gene induction in a GAL3 mutant of Saccharomyces cerevisiae (pages 1798–1817)

      Rajesh Kumar Kar, Mohd. Tanvir Qureshi, Akshay Kumar DasAdhikari, Taiyeb Zahir, Kareenhalli V. Venkatesh and Paike Jayadeva Bhat

      Article first published online: 28 FEB 2014 | DOI: 10.1111/febs.12741

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      Unlike a wild-type strain, gal3∆ strain shows long-term adaptation towards galactose. Using growth complementation assays, flow cytometry and microscopic analysis we demonstrate that only 0.3% of gal3∆ cell population respond to galactose and which is fixed. The induced and un-induced population can switch back to their original bimodal state. The gal3∆ cells activate the GAL pathway by implementing non-genetic heterogeneity.

    8. Structure, diversity and evolution of myriapod hemocyanins (pages 1818–1833)

      Christian Pick, Samantha Scherbaum, Elöd Hegedüs, Andreas Meyer, Michael Saur, Ruben Neumann, Jürgen Markl and Thorsten Burmester

      Article first published online: 5 MAR 2014 | DOI: 10.1111/febs.12742

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      Hemocyanin transports O2 in many arthropods. We demonstrate a widespread occurrence hemocyanin in centipedes, millipedes and symphylans. Some species lack hemocyanin. Two distinct subunit types evolved early in myriapod evolution. We further show the presence of hemocyanin-like phenoloxidases in this taxon. Specifically, the structure of the 3 × 6-mer hemocyanin of Polydesmus angustus was revealed by sequencing, 3D electron microscopy and homology modelling.

    9. Thermodynamic and structural analysis of HIV protease resistance to darunavir – analysis of heavily mutated patient-derived HIV-1 proteases (pages 1834–1847)

      Milan Kožíšek, Martin Lepšík, Klára Grantz Šašková, Jiří Brynda, Jan Konvalinka and Pavlína Řezáčová

      Article first published online: 28 FEB 2014 | DOI: 10.1111/febs.12743

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      We report enzymologic, thermodynamic and structural analyses of a series of six clinically derived mutant HIV proteases (PR) resistant to darunavir. We show that even very dramatic changes in PR sequence leading to the loss of hydrogen bonds with the inhibitor could be partially compensated by the entropy contribution due to burial of larger nonpolar surface area of the protein.

    10. Investigations into the auto-FAT10ylation of the bispecific E2 conjugating enzyme UBA6-specific E2 enzyme 1 (pages 1848–1859)

      Annette Aichem, Nicola Catone and Marcus Groettrup

      Article first published online: 3 MAR 2014 | DOI: 10.1111/febs.12745

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      The ubiquitin-like modifier FAT10 is conjugated to its substrates via the bispecific, ubiquitin- and FAT10-activating enzyme UBA6, the bi-specific conjugating enzyme USE1, and possibly E3 ligases. USE1 auto-FAT10ylation leads to its proteasomal degradation and serves as a negative feedback mechanism to limit the FAT10 conjugation pathway. Interestingly, it does not inactivate USE1 concerning ubiquitin or FAT10 conjugation.

    11. Homooligomerization is needed for stability: a molecular modelling and solution study of Escherichia coli purine nucleoside phosphorylase (pages 1860–1871)

      Branimir Bertoša, Goran Mikleušević, Beata Wielgus-Kutrowska, Marta Narczyk, Matea Hajnić, Ivana Leščić Ašler, Sanja Tomić, Marija Luić and Agnieszka Bzowska

      Article first published online: 3 MAR 2014 | DOI: 10.1111/febs.12746

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      Although the dimer seems as sufficient form for conducting catalysis, E. coli purine nucleoside phosphorylase exists as a hexamer composed as trimer of dimers. We investigated structural and dynamical properties of the enzyme, and some dimer-dimer interface mutants, and concluded that the hexameric architecture is necessary since it provides the stabilization of the proper three-dimensional structure of the dimeric assembly.

    12. MicroRNA expression and regulation in the uterus during embryo implantation in rat (pages 1872–1891)

      Hong-Fei Xia, Xiao-Hua Jin, Zong-Fu Cao, Yi Hu and Xu Ma

      Article first published online: 7 MAR 2014 | DOI: 10.1111/febs.12751

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      MiR-29a was highly expressed in rat uterus during the implantation period. Both pro-apoptotic (Bak1 and Bmf) and anti-apoptotic factors (Bcl-w) were the target genes of miR-29a. There was weak binding capacity between miR-29a and 3′-UTR of anti-apoptotic factor Mcl1. Over-expression of miR-29a inhibited the late apoptosis of endometrial stromal cells, which may be caused by predominately combining pro-apoptotic factors.

    13. Role of activating transcription factor 3 protein ATF3 in necrosis and apoptosis induced by 5-fluoro-2′-deoxyuridine (pages 1892–1900)

      Akira Sato, Kentaro Nakama, Hiroki Watanabe, Akito Satake, Akihiro Yamamoto, Takuya Omi, Akiko Hiramoto, Mitsuko Masutani, Yusuke Wataya and Hye-Sook Kim

      Article first published online: 6 MAR 2014 | DOI: 10.1111/febs.12752

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      We have analyzed the mechanisms regulating the necrosis and the apoptosis that occur on treatment of mouse cancer FM3A cells with antitumor FUdR. The transcription factor ATF3 was induced strongly in the necrosis. Knockdown of Atf3 in the necrosis-fated cells caused apoptosis instead of necrosis. These findings suggest that the ATF3 is a regulator in the necrosis and the apoptosis.

    14. Glucose de-repression by yeast AMP-activated protein kinase SNF1 is controlled via at least two independent steps (pages 1901–1917)

      Raúl García-Salcedo, Timo Lubitz, Gemma Beltran, Karin Elbing, Ye Tian, Simone Frey, Olaf Wolkenhauer, Marcus Krantz, Edda Klipp and Stefan Hohmann

      Article first published online: 4 MAR 2014 | DOI: 10.1111/febs.12753

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      The AMP-activated protein kinase, AMPK, controls energy homeostasis in eukaryotic cells. Yeast AMPK, SNF1, mediates glucose de-repression. Here we employed time course experimentation and modelling to show that glucose de-repression via SNF1 is regulated by glucose via at least two independent steps: the control of activation of the Snf1 kinase and directing active Snf1 to its target Mig1.

    15. Molecular characterization of ubiquitin-specific protease 18 reveals substrate specificity for interferon-stimulated gene 15 (pages 1918–1928)

      Anja Basters, Paul P. Geurink, Farid El Oualid, Lars Ketscher, Marco S. Casutt, Eberhard Krause, Huib Ovaa, Klaus-Peter Knobeloch and Günter Fritz

      Article first published online: 3 MAR 2014 | DOI: 10.1111/febs.12754

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      Protein modification by the interferon-stimulated gene 15 (ISG15), an ubiquitin-like modifier, represents one of the major antiviral effector systems. Covalent linkage of ISG15 to proteins is counteracted by the protease USP18. We established expression and purification of USP18 from insect cells and show that the enzyme is highly specific for ISG15, cleaving ISG15 but not ubiquitin from different substrates.

  5. Author index

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. You have free access to this content
      Author index (page 1929)

      Article first published online: 1 APR 2014 | DOI: 10.1111/febs.12646

  6. Table of Contents

    1. Top of page
    2. Front Cover
    3. Editorial Information
    4. Editor's Choice
    5. Original Articles
    6. Author index
    7. Table of Contents
    1. Table of Contents (page 1930)

      Article first published online: 1 APR 2014 | DOI: 10.1111/febs.12647

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