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https://www.readbyqxmd.com/read/27920294/effects-of-increasing-the-affinity-of-card-for-rna-polymerase-on-mycobacterium-tuberculosis-growth-rrna-transcription-and-virulence
#1
Ashley L Garner, Jayan Rammohan, Jeremy P Huynh, Lucas M Onder, James Chen, Brian Bae, Drake Jensen, Leslie A Weiss, Ana Ruiz Manzano, Seth A Darst, Elizabeth A Campbell, Bryce E Nickels, Eric A Galburt, Christina L Stallings
: CarD is an essential RNA polymerase (RNAP) interacting protein in Mycobacterium tuberculosis (Mtb) that stimulates formation of RNAP-promoter open complexes. CarD plays a complex role in Mtb growth and virulence that is not fully understood. Therefore, to gain further insight into the role of CarD in Mtb growth and virulence we determined the effect of increasing the affinity of CarD for RNAP. Using site-directed mutagenesis guided by crystal structures of CarD bound to RNAP, we identified amino acid substitutions that increase the affinity of CarD for RNAP...
December 5, 2016: Journal of Bacteriology
https://www.readbyqxmd.com/read/27906650/structure-of-the-70s-ribosome-from-human-pathogen-staphylococcus-aureus
#2
Iskander Khusainov, Quentin Vicens, Anthony Bochler, François Grosse, Alexander Myasnikov, Jean-François Ménétret, Johana Chicher, Stefano Marzi, Pascale Romby, Gulnara Yusupova, Marat Yusupov, Yaser Hashem
Comparative structural studies of ribosomes from various organisms keep offering exciting insights on how species-specific or environment-related structural features of ribosomes may impact translation specificity and its regulation. Although the importance of such features may be less obvious within more closely related organisms, their existence could account for vital yet species-specific mechanisms of translation regulation that would involve stalling, cell survival and antibiotic resistance. Here, we present the first full 70S ribosome structure from Staphylococcus aureus, a Gram-positive pathogenic bacterium, solved by cryo-electron microscopy...
December 1, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27906161/structural-basis-for-arfa-rf2-mediated-translation-termination-on-mrnas-lacking-stop-codons
#3
Paul Huter, Claudia Müller, Bertrand Beckert, Stefan Arenz, Otto Berninghausen, Roland Beckmann, Daniel N Wilson
In bacteria, ribosomes stalled on truncated mRNAs lacking a stop codon are rescued by either the transfer-messenger RNA (tmRNA), alternative rescue factor A (ArfA) or ArfB rescue systems(1). While tmRNA- and ArfB-ribosome structures have been elucidated(2-7), structural insight into how ArfA recognizes the presence of truncated mRNAs and recruits the canonical termination release factor RF2 to rescue the stalled ribosomes is lacking. Here we present a cryo-electron microscopy reconstruction of a ribosome stalled on a truncated mRNA in the presence of ArfA and RF2...
December 1, 2016: Nature
https://www.readbyqxmd.com/read/27906160/mechanistic-insights-into-the-alternative-translation-termination-by-arfa-and-rf2
#4
Chengying Ma, Daisuke Kurita, Ningning Li, Yan Chen, Hyouta Himeno, Ning Gao
During cellular translation of mRNAs by ribosomes, various situations that would result in the pausing or stalling of translation apparatus at elongation and termination steps often occur(1-6). Except for programmed stalling, which is usually utilized by the cells to exert regulatory purposes(5,7,8), ribosomes stalled on mRNAs need to be terminated and recycled to maintain adequate cellular translation capacity(9). A large source of ribosome stalling originates in aberrant mRNAs that lack a stop codon. Transcriptional errors and misprocessing of primary transcripts, as well as undesired mRNA cleavage, all contribute to the formation of non-stop mRNAs...
December 1, 2016: Nature
https://www.readbyqxmd.com/read/27892503/trans-translation-is-essential-in-the-human-pathogen-legionella-pneumophila
#5
Romain Brunel, Xavier Charpentier
Trans-translation is a ubiquitous bacterial mechanism for ribosome rescue in the event of translation stalling. Although trans-translation is not essential in several bacterial species, it has been found essential for viability or virulence in a wide range of pathogens. We describe here that trans-translation is essential in the human pathogen Legionella pneumophila, the etiologic agent of Legionnaire's disease (LD), a severe form of nosocomial and community-acquired pneumonia. The ssrA gene coding for tmRNA, the key component of trans-translation, could not be deleted in L...
November 28, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27829146/absence-of-non-histone-protein-complexes-at-natural-chromosomal-pause-sites-results-in-reduced-replication-pausing-in-aging-yeast-cells
#6
Marleny Cabral, Xin Cheng, Sukhwinder Singh, Andreas S Ivessa
There is substantial evidence that genomic instability increases during aging. Replication pausing (and stalling) at difficult-to-replicate chromosomal sites may induce genomic instability. Interestingly, in aging yeast cells, we observed reduced replication pausing at various natural replication pause sites (RPSs) in ribosomal DNA (rDNA) and non-rDNA locations (e.g., silent replication origins and tRNA genes). The reduced pausing occurs independent of the DNA helicase Rrm3p, which facilitates replication past these non-histone protein-complex-bound RPSs, and is independent of the deacetylase Sir2p...
November 8, 2016: Cell Reports
https://www.readbyqxmd.com/read/27827794/molecular-insights-into-protein-synthesis-with-proline-residues
#7
Sergey Melnikov, Justine Mailliot, Lukas Rigger, Sandro Neuner, Byung-Sik Shin, Gulnara Yusupova, Thomas E Dever, Ronald Micura, Marat Yusupov
Proline is an amino acid with a unique cyclic structure that facilitates the folding of many proteins, but also impedes the rate of peptide bond formation by the ribosome. As a ribosome substrate, proline reacts markedly slower when compared with other amino acids both as a donor and as an acceptor of the nascent peptide. Furthermore, synthesis of peptides with consecutive proline residues triggers ribosome stalling. Here, we report crystal structures of the eukaryotic ribosome bound to analogs of mono- and diprolyl-tRNAs...
December 2016: EMBO Reports
https://www.readbyqxmd.com/read/27821493/mechanistic-insights-into-mammalian-stress-granule-dynamics
#8
REVIEW
Marc D Panas, Pavel Ivanov, Paul Anderson
The accumulation of stalled translation preinitiation complexes (PICs) mediates the condensation of stress granules (SGs). Interactions between prion-related domains and intrinsically disordered protein regions found in SG-nucleating proteins promote the condensation of ribonucleoproteins into SGs. We propose that PIC components, especially 40S ribosomes and mRNA, recruit nucleators that trigger SG condensation. With resolution of stress, translation reinitiation reverses this process and SGs disassemble. By cooperatively modulating the assembly and disassembly of SGs, ribonucleoprotein condensation can influence the survival and recovery of cells exposed to unfavorable environmental conditions...
November 7, 2016: Journal of Cell Biology
https://www.readbyqxmd.com/read/27791127/cockayne-syndrome-group-a-and-b-proteins-converge-on-transcription-linked-resolution-of-non-b-dna
#9
Morten Scheibye-Knudsen, Anne Tseng, Martin Borch Jensen, Karsten Scheibye-Alsing, Evandro Fei Fang, Teruaki Iyama, Sanjay Kumar Bharti, Krisztina Marosi, Lynn Froetscher, Henok Kassahun, David Mark Eckley, Robert W Maul, Paul Bastian, Supriyo De, Soumita Ghosh, Hilde Nilsen, Ilya G Goldberg, Mark P Mattson, David M Wilson, Robert M Brosh, Myriam Gorospe, Vilhelm A Bohr
Cockayne syndrome is a neurodegenerative accelerated aging disorder caused by mutations in the CSA or CSB genes. Although the pathogenesis of Cockayne syndrome has remained elusive, recent work implicates mitochondrial dysfunction in the disease progression. Here, we present evidence that loss of CSA or CSB in a neuroblastoma cell line converges on mitochondrial dysfunction caused by defects in ribosomal DNA transcription and activation of the DNA damage sensor poly-ADP ribose polymerase 1 (PARP1). Indeed, inhibition of ribosomal DNA transcription leads to mitochondrial dysfunction in a number of cell lines...
November 1, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27791002/context-specific-inhibition-of-translation-by-ribosomal-antibiotics-targeting-the-peptidyl-transferase-center
#10
James Marks, Krishna Kannan, Emily J Roncase, Dorota Klepacki, Amira Kefi, Cédric Orelle, Nora Vázquez-Laslop, Alexander S Mankin
The first broad-spectrum antibiotic chloramphenicol and one of the newest clinically important antibacterials, linezolid, inhibit protein synthesis by targeting the peptidyl transferase center of the bacterial ribosome. Because antibiotic binding should prevent the placement of aminoacyl-tRNA in the catalytic site, it is commonly assumed that these drugs are universal inhibitors of peptidyl transfer and should readily block the formation of every peptide bond. However, our in vitro experiments showed that chloramphenicol and linezolid stall ribosomes at specific mRNA locations...
October 25, 2016: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/27784783/the-ribosome-restrains-molten-globule-formation-in-stalled-nascent-flavodoxin
#11
Joseline A Houwman, Estelle André, Adrie H Westphal, Willem J H van Berkel, Carlo P M van Mierlo
Folding of proteins usually involves intermediates, of which an important type is the molten globule (MG). MGs are ensembles of interconverting conformers that contain (non-)native secondary structure and lack the tightly packed tertiary structure of natively folded globular proteins. Whereas MGs of various purified proteins have been probed to date, no data is available on their presence and/or effect during protein synthesis. To study whether MGs arise during translation, we use ribosome-nascent chain (RNC) complexes of the electron-transfer protein flavodoxin...
October 26, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27760805/the-minimum-open-reading-frame-aug-stop-induces-boron-dependent-ribosome-stalling-and-mrna-degradation
#12
Mayuki Tanaka, Naoyuki Sotta, Yusuke Yamazumi, Yui Yamashita, Kyoko Miwa, Katsunori Murota, Yukako Chiba, Masami Yokota Hirai, Tetsu Akiyama, Hitoshi Onouchi, Satoshi Naito, Toru Fujiwara
Upstream open reading frames (uORFs) are often translated ahead of the main ORF of a gene and regulate gene expression, sometimes in a condition-dependent manner, but such a role for the minimum uORF (hereafter referred to as 'AUG-stop') in living organisms is currently unclear. Here, we show that AUG-stop plays an important role in the boron (B)-dependent regulation of NIP5;1, encoding a boric acid channel required for normal growth under low B conditions in Arabidopsis thaliana. High B enhanced ribosome stalling at AUG-stop, which was accompanied by the suppression of translation and mRNA degradation...
October 19, 2016: Plant Cell
https://www.readbyqxmd.com/read/27758893/genome-wide-mapping-of-uncapped-and-cleaved-transcripts-reveals-a-role-for-the-nuclear-mrna-cap-binding-complex-in-cotranslational-rna-decay-in-arabidopsis
#13
Xiang Yu, Matthew R Willmann, Stephen J Anderson, Brian D Gregory
RNA turnover is necessary for controlling proper mRNA levels posttranscriptionally. In general, RNA degradation is via exoribonucleases that degrade RNA either from the 5' end to the 3' end, such as XRN4, or in the opposite direction by the multisubunit exosome complex. Here, we use genome-wide mapping of uncapped and cleaved transcripts to reveal the global landscape of cotranslational mRNA decay in the Arabidopsis thaliana transcriptome. We found that this process leaves a clear three nucleotide periodicity in open reading frames...
October 2016: Plant Cell
https://www.readbyqxmd.com/read/27742800/global-analysis-of-truncated-rna-ends-reveals-new-insights-into-ribosome-stalling-in-plants
#14
Cheng-Yu Hou, Wen-Chi Lee, Hsiao-Chun Chou, Ai-Ping Chen, Shu-Jen Chou, Ho-Ming Chen
High-throughput approaches for profiling the 5' ends of RNA degradation intermediates on a genome-wide scale are frequently applied to analyze and validate cleavage sites guided by microRNAs (miRNAs). However, the complexity of the RNA degradome other than miRNA targets is currently largely uncharacterized, and this limits the application of RNA degradome studies. We conducted a global analysis of 5'-truncated mRNA ends that mapped to coding sequences (CDSs) of Arabidopsis thaliana, rice (Oryza sativa), and soybean (Glycine max)...
October 2016: Plant Cell
https://www.readbyqxmd.com/read/27736765/sorting-out-antibiotics-mechanisms-of-action-a-double-fluorescent-protein-reporter-for-high-throughput-screening-of-ribosome-and-dna-biosynthesis-inhibitors
#15
Ilya A Osterman, Ekaterina S Komarova, Dmitry I Shiryaev, Ilya A Korniltsev, Irina M Khven, Dmitry A Lukyanov, Vadim N Tashlitsky, Marina V Serebryakova, Olga V Efremenkova, Yan A Ivanenkov, Alexey A Bogdanov, Petr V Sergiev, Olga A Dontsova
In order to accelerate drug discovery, a simple, reliable, and cost-effective system for high-throughput identification of a potential antibiotic mechanism of action is required. To facilitate such screening of new antibiotics, we created a double-reporter system for not only antimicrobial activity detection but also simultaneous sorting of potential antimicrobials into those that cause ribosome stalling and those that induce the SOS response due to DNA damage. In this reporter system, the red fluorescent protein gene rfp was placed under the control of the SOS-inducible sulA promoter...
December 2016: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/27677373/the-aaa-ftsh-protease-degrades-an-ssra-tagged-model-protein-in-the-inner-membrane-of-escherichia-coli
#16
Sanjay B Hari, Robert T Sauer
In eubacteria, the tmRNA system frees ribosomes that stall during protein synthesis and adds an ssrA tag to the incompletely translated polypeptide to target it for degradation. The AAA+ ClpXP protease degrades most ssrA-tagged proteins in the Escherichia coli cytoplasm and was recently shown to degrade an ssrA-tagged protein in the inner membrane. However, we find that tmRNA-mediated tagging of E. coli ProW1-182, a different inner-membrane protein, results in degradation by the membrane-tethered AAA+ FtsH protease...
October 11, 2016: Biochemistry
https://www.readbyqxmd.com/read/27645242/the-expression-of-antibiotic-resistance-methyltransferase-correlates-with-mrna-stability-independently-of-ribosome-stalling
#17
Ekaterina Dzyubak, M N Yap
Members of the Erm methyltransferase family modify 23S rRNA of the bacterial ribosome and render cross-resistance to macrolides and multiple distantly related antibiotics. Previous studies have shown that the expression of erm is activated when a macrolide-bound ribosome stalls the translation of the leader peptide preceding the cotranscribed erm Ribosome stalling is thought to destabilize the inhibitory stem-loop mRNA structure and exposes the erm Shine-Dalgarno (SD) sequence for translational initiation. Paradoxically, mutations that abolish ribosome stalling are routinely found in hyper-resistant clinical isolates; however, the significance of the stalling-dead leader sequence is largely unknown...
December 2016: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/27624127/maintenance-of-transcription-translation-coupling-by-elongation-factor-p
#18
Sara Elgamal, Irina Artsimovitch, Michael Ibba
UNLABELLED: Under conditions of tight coupling between translation and transcription, the ribosome enables synthesis of full-length mRNAs by preventing both formation of intrinsic terminator hairpins and loading of the transcription termination factor Rho. While previous studies have focused on transcription factors, we investigated the role of Escherichia coli elongation factor P (EF-P), an elongation factor required for efficient translation of mRNAs containing consecutive proline codons, in maintaining coupled translation and transcription...
2016: MBio
https://www.readbyqxmd.com/read/27588015/genetic-selection-of-peptide-aptamers-that-interact-and-inhibit-both-small-protein-b-and-alternative-ribosome-rescue-factor-a-of-aeromonas-veronii-c4
#19
Peng Liu, Yong Chen, Dan Wang, Yanqiong Tang, Hongqian Tang, Haichao Song, Qun Sun, Yueling Zhang, Zhu Liu
Aeromonas veronii is a pathogenic gram-negative bacterium, which infects a variety of animals and results in mass mortality. The stalled-ribosome rescues are reported to ensure viability and virulence under stress conditions, of which primarily include trans-translation and alternative ribosome-rescue factor A (ArfA) in A. veronii. For identification of specific peptides that interact and inhibit the stalled-ribosome rescues, peptide aptamer library (pTRG-SN-peptides) was constructed using pTRG as vector and Staphylococcus aureus nuclease (SN) as scaffold protein, in which 16 random amino acids were introduced to form an exposed surface loop...
2016: Frontiers in Microbiology
https://www.readbyqxmd.com/read/27551685/structural-analysis-of-a-signal-peptide-inside-the-ribosome-tunnel-by-dnp-mas-nmr
#20
Sascha Lange, W Trent Franks, Nandhakishore Rajagopalan, Kristina Döring, Michel A Geiger, Arne Linden, Barth-Jan van Rossum, Günter Kramer, Bernd Bukau, Hartmut Oschkinat
Proteins are synthesized in cells by ribosomes and, in parallel, prepared for folding or targeting. While ribosomal protein synthesis is progressing, the nascent chain exposes amino-terminal signal sequences or transmembrane domains that mediate interactions with specific interaction partners, such as the signal recognition particle (SRP), the SecA-adenosine triphosphatase, or the trigger factor. These binding events can set the course for folding in the cytoplasm and translocation across or insertion into membranes...
August 2016: Science Advances
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