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Engineering RNA endonucleases with customized sequence specificities
Sequence-specific DNA endonucleases have found numerous applications in biology, but similar manipulations of RNA have been limited by the lack of suitable enzymes. These authors combine a cleavage domain with a designable binding domain and demonstrate the resulting RNA endonuclease's utilityin vitroand in cells.
- Rajarshi Choudhury
- , Yihsuan S. Tsai
- & Zefeng Wang
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Lysine methylation of VCP by a member of a novel human protein methyltransferase family
Methyltransferases modify cellular proteins in addition to DNA and histones. These authors identify a new family of lysine-specific methyltransferases and show that a member of this family, which is associated with tumour metastasis, methylates the ATP-dependent protein chaperone VCP/p97.
- Stefan Kernstock
- , Erna Davydova
- & Pål Ø. Falnes
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| Open AccessPKA regulatory subunits mediate synergy among conserved G-protein-coupled receptor cascades
G-protein-coupled receptors sense extracellular cues and transmit the signal to distinct trimeric G-proteins. Stefanet al.show that in response to cAMP, a central and conserved component of the Gαs-coupled receptor cascade, the RII subunit of PKA, specifically binds to and participates in Gαi signaling.
- Eduard Stefan
- , Mohan K. Malleshaiah
- & Stephen W. Michnick
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Synapse microarray identification of small molecules that enhance synaptogenesis
Large scale synapse assays can facilitate identification of drug leads. Shiet al. develop a 'synapse microarray' technology that enables sensitive, high-throughput, quantitative screening of synaptogenic events, and use it to identify novel histone deacetylase inhibitors that enhance synaptogenesis.
- Peng Shi
- , Mark A. Scott
- & Mehmet Fatih Yanik
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Crystal structure of the human thioredoxin reductase–thioredoxin complex
Thioredoxin reductase 1 is a flavoprotein that is involved in the regulation of redox homeostasis. Fritz-Wolfet al. report the first crystal structures of thioredoxin reductase 1 in complex with its substrate, thioredoxin, and confirm that the enzyme uses a flexible carboxy-terminal arm for electron transport to its substrates.
- Karin Fritz-Wolf
- , Sebastian Kehr
- & Katja Becker
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Identification and characterization of a multidomain hyperthermophilic cellulase from an archaeal enrichment
Archaea are microorganisms that use a wide range of carbon and energy sources. Grahamet al. describe an archaeal consortium that can grow at temperatures above 90 °C using crystalline cellulose as a carbon source, with potential applications in enzymatic degradation under extreme conditions.
- Joel E. Graham
- , Melinda E. Clark
- & Frank T. Robb
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Potential for interdependent development of tRNA determinants for aminoacylation and ribosome decoding
Aminoacyl-transfer RNA synthetases are conserved between bacteria and eukaryotes; however, bacterial enzymes cannot acylate eukaryote tRNAs. Now, fusion of a human and bacterial enzyme is shown to overcome the species barrier and confer tRNA specificity during both codon selection and proofreading on the ribosome.
- Cuiping Liu
- , Howard Gamper
- & Ya-Ming Hou
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| Open AccessPredicting sites of ADAR editing in double-stranded RNA
ADAR enzymes edit double-stranded RNA, converting adenosines to inosines, and are essential for neuronal function. Eggingtonet al. quantify edit sites in RNA using a Sanger sequencing protocol and use the resulting data to develop algorithms to predict RNA edit sites.
- Julie M. Eggington
- , Tom Greene
- & Brenda L. Bass
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| Open AccessPhotosynthetic growth despite a broken Q-cycle
The Q-cycle is thought to be an essential energetic component of the photosynthetic electron-transfer chain. Here, Chlamydomonas mutants with an inactive Q-cycle but normal levels ofb6fcomplexes are shown to display photosynthetic growth, demonstrating the dispensability of the Q-cycle in the oxygenic photosynthetic chain.
- Alizée Malnoë
- , Francis-André Wollman
- & Fabrice Rappaport
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Two-dimensional enzyme diffusion in laterally confined DNA monolayers
Little is known about the effects of molecular crowding and confinement on biomolecule function. Castronovoet al. investigate the reactions of restriction enzymes with DNA confined in bushy matrices and find that the enzymes enter at the side of the matrix before diffusing two-dimensionally.
- Matteo Castronovo
- , Agnese Lucesoli
- & Giacinto Scoles
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ALKBH8-mediated formation of a novel diastereomeric pair of wobble nucleosides in mammalian tRNA
Uridines at the wobble position of transfer RNA anticodons are usually modified to allow efficient decoding of messenger RNA codons. In this study, ALKBH8 is shown to be a bifunctional transfer RNA modification enzyme required for the formation of a novel diastereomeric pair of modified wobble uridines.
- Erwin van den Born
- , Cathrine B. Vågbø
- & Pål Ø. Falnes
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The phospholipase D1 pathway modulates macroautophagy
Macroautophagy involves remodelling of intracellular membranes, but the role of lipids and lipid enzymes in this process is poorly understood. Here, the authors demonstrate that phospholipase D1, an enzyme that catalyses the production of phosphatidic acid, has a modulatory role in the autophagy process in mammalian cells.
- Claudia Dall'Armi
- , Andrés Hurtado-Lorenzo
- & Gilbert Di Paolo
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High rates of photobiological H2 production by a cyanobacterium under aerobic conditions
Hydrogen production using photosynthetic bacteria is an appealing energy source, but typically the bacteria require anaerobic conditions. Here, the authors report a wild-type cyanobacterium strain that shows very high rates of hydrogen production under aerobic environmental conditions.
- Anindita Bandyopadhyay
- , Jana Stöckel
- & Himadri B. Pakrasi
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| Open AccessSubstrate docking to γ-secretase allows access of γ-secretase modulators to an allosteric site
γ-Secretase modulators have promise in the treatment of Alzheimer's disease, but their molecular target is uncertain. Here, fluorescence resonance energy transfer is used to determine that the γ-secretase allosteric site is within the γ-secretase complex and that substrate docking is required for modulators to access the site.
- Kengo Uemura
- , Katherine C. Farner
- & Oksana Berezovska
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Overlap between folding and functional energy landscapes for adenylate kinase conformational change
Enzyme function is often dependent on fluctuations between inactive and active states. Olsson and Wolf-Watz show that switching between the inactive and active states of adenylate kinase is associated with partial unfolding/refolding of the enzyme.
- Ulrika Olsson
- & Magnus Wolf-Watz
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Development of a novel selective inhibitor of the Down syndrome-related kinase Dyrk1A
The kinase Dyrk1A is essential for brain function and development, and its excessive activity has been implicated in Down syndrome. In this study, a selective inhibitor of Dyrk1A is developed, which may help to elucidate the molecular mechanisms of normal and diseased brain.
- Yasushi Ogawa
- , Yosuke Nonaka
- & Masatoshi Hagiwara