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https://www.readbyqxmd.com/read/28202229/requirement-of-the-rna-binding-protein-smpb-during-intracellular-growth-of-listeria-monocytogenes
#1
Mobarak Abu Mraheil, Renate Frantz, Lisa Teubner, Heiko Wendt, Uwe Linne, Jessica Wingerath, Thomas Wirth, Trinad Chakraborty
Bacterial trans-translation is the main quality control mechanism employed to relieve stalled ribosomes. Trans-translation is mediated by the small protein B (SmpB) and transfer-mRNA (tmRNA) ribonucleoprotein complex, which interacts with translational complexes stalled at the 3' end of non-stop mRNAs to release the stalled ribosomes thereby targeting the nascent polypeptides and truncated mRNAs for degradation. The trans-translation system exists with a few exceptions in all bacteria. In the present study, we assessed the contribution of SmpB to the growth and virulence of Listeria monocytogenes, a human intracellular food-borne pathogen that colonizes host tissues to cause severe invasive infections...
February 1, 2017: International Journal of Medical Microbiology: IJMM
https://www.readbyqxmd.com/read/28193672/translation-of-poly-a-tails-leads-to-precise-mrna-cleavage
#2
Nicholas R Guydosh, Rachel Green
Translation of poly(A) tails leads to mRNA cleavage but the mechanism and global pervasiveness of this "nonstop/no-go" decay process is not understood. Here we performed ribosome profiling (in a yeast strain lacking exosome function) of short 15-18 nt mRNA footprints to identify ribosomes stalled at 3' ends of mRNA decay intermediates. In this background, we found mRNA cleavage extending hundreds of nucleotides upstream of ribosome stalling in poly(A) and predominantly in one reading frame. These observations suggest that decay-triggering endonucleolytic cleavage is closely associated with the ribosome...
February 13, 2017: RNA
https://www.readbyqxmd.com/read/28181542/elongation-factor-p-restricts-salmonella-s-growth-by-controlling-translation-of-a-mg-2-transporter-gene-during-infection
#3
Eunna Choi, Soomin Choi, Daesil Nam, Shinae Park, Yoontak Han, Jung-Shin Lee, Eun-Jin Lee
When a ribosome translates mRNA sequences, the ribosome often stalls at certain codons because it is hard to translate. Consecutive proline codons are such examples that induce ribosome stalling and elongation factor P (EF-P) is required for the stalled ribosome to continue translation at those consecutive proline codons. We found that EF-P is required for translation of the mgtB gene encoding a Mg(2+) transporter in the mgtCBR virulence operon from the intracellular pathogen Salmonella enterica serovar Typhimurium...
February 9, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28138069/ribosome-pausing-arrest-and-rescue-in-bacteria-and-eukaryotes
#4
REVIEW
Allen R Buskirk, Rachel Green
Ribosomes translate genetic information into polypeptides in several basic steps: initiation, elongation, termination and recycling. When ribosomes are arrested during elongation or termination, the cell's capacity for protein synthesis is reduced. There are numerous quality control systems in place to distinguish between paused ribosomes that need some extra input to proceed and terminally stalled ribosomes that need to be rescued. Here, we discuss similarities and differences in the systems for resolution of pauses and rescue of arrested ribosomes in bacteria and eukaryotes, and how ribosome profiling has transformed our ability to decipher these molecular events...
March 19, 2017: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
https://www.readbyqxmd.com/read/28132843/znf598-and-rack1-regulate-mammalian-ribosome-associated-quality-control-function-by-mediating-regulatory-40s-ribosomal-ubiquitylation
#5
Elayanambi Sundaramoorthy, Marilyn Leonard, Raymond Mak, Jeffrey Liao, Amitkumar Fulzele, Eric J Bennett
Ribosomes that experience terminal stalls during translation are resolved by ribosome-associated quality control (QC) pathways that oversee mRNA and nascent chain destruction and recycle ribosomal subunits. The proximal factors that sense stalled ribosomes and initiate mammalian ribosome-associated QC events remain undefined. We demonstrate that the ZNF598 ubiquitin ligase and the 40S ribosomal protein RACK1 help to resolve poly(A)-induced stalled ribosomes. They accomplish this by regulating distinct and overlapping regulatory 40S ribosomal ubiquitylation events...
January 26, 2017: Molecular Cell
https://www.readbyqxmd.com/read/28115162/nonstop-mrna-decay-machinery-is-involved-in-the-clearance-of-mrna-5-fragments-produced-by-rnai-and-nmd-in-drosophila-melanogaster-cells
#6
Yoshifumi Hashimoto, Masaki Takahashi, Eri Sakota, Yoshikazu Nakamura
When translating mRNAs are cleaved in protein-coding regions, 5' fragments of mRNAs are detached from stop codons (i.e., nonstop mRNAs) and protected from 3'-5' exonucleases by ribosomes stalled at the 3' termini. It has been shown in yeast that the nonstop mRNA decay (NSD) machinery triggers nonstop mRNA degradation by removing stalled ribosomes in the artificial reporter mRNAs. However, it is not known well whether NSD is involved in the degradation of endogenous nonstop mRNAs in higher eukaryotes. In this work, we addressed the question of whether 5'-nonstop-mRNA fragments generated by siRNA cleavage or nonsense-mediated-mRNA decay (NMD) are degraded by the NSD pathway in Drosophila melanogaster cells by knocking down three NSD components, Pelota (a yeast Dom34 homolog), Hbs1 and ABCE1 (a ribosome-recycling factor)...
January 20, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28113053/mazef-toxin-antitoxin-proteins-alter-e-coli-cell-morphology-and-infrastructure-during-persister-formation-and-regrowth
#7
Junho Cho, Anita Carr, Lisa Whitworth, Brent Johnson, Kevin Scott Wilson
When exposed to antibiotics, many bacteria respond by activating intracellular "toxin" proteins, which arrest cell growth and induce formation of persister cells that survive antibiotics. After antibiotics are removed, persisters can regrow by synthesizing "antitoxin" proteins that sequester toxin proteins. In E. coli, MazE antitoxin sequesters the activity of MazF toxin, which extensively cleaves cellular RNAs. Although the functions of MazEF proteins are well characterized, there is surprisingly little known about their effects on cell structure...
January 22, 2017: Microbiology
https://www.readbyqxmd.com/read/28077875/structural-basis-of-co-translational-quality-control-by-arfa-and-rf2-bound-to-ribosome
#8
Fuxing Zeng, Yanbo Chen, Jonathan Remis, Mrinal Shekhar, James C Phillips, Emad Tajkhorshid, Hong Jin
Quality control mechanisms intervene appropriately when defective translation events occur, in order to preserve the integrity of protein synthesis. Rescue of ribosomes translating on messenger RNAs that lack stop codons is one of the co-translational quality control pathways. In many bacteria, ArfA recognizes stalled ribosomes and recruits the release factor RF2, which catalyses the termination of protein synthesis. Although an induced-fit mechanism of nonstop mRNA surveillance mediated by ArfA and RF2 has been reported, the molecular interaction between ArfA and RF2 in the ribosome that is responsible for the mechanism is unknown...
January 26, 2017: Nature
https://www.readbyqxmd.com/read/28065601/initiation-of-quality-control-during-poly-a-translation-requires-site-specific-ribosome-ubiquitination
#9
Szymon Juszkiewicz, Ramanujan S Hegde
Diverse cellular stressors have been observed to trigger site-specific ubiquitination on several ribosomal proteins. However, the ubiquitin ligases, biochemical consequences, and physiologic pathways linked to these modifications are not known. Here, we show in mammalian cells that the ubiquitin ligase ZNF598 is required for ribosomes to terminally stall during translation of poly(A) sequences. ZNF598-mediated stalling initiated the ribosome-associated quality control (RQC) pathway for degradation of nascent truncated proteins...
December 22, 2016: Molecular Cell
https://www.readbyqxmd.com/read/28008922/atp-hydrolysis-by-upf1-is-required-for-efficient-translation-termination-at-premature-stop-codons
#10
Lucas D Serdar, DaJuan L Whiteside, Kristian E Baker
Nonsense-mediated mRNA decay (NMD) represents a eukaryotic quality control pathway that recognizes and rapidly degrades transcripts harbouring nonsense mutations to limit accumulation of non-functional and potentially toxic truncated polypeptides. A critical component of the NMD machinery is UPF1, an RNA helicase whose ATPase activity is essential for NMD, but for which the precise function and site of action remain unclear. We provide evidence that ATP hydrolysis by UPF1 is required for efficient translation termination and ribosome release at a premature termination codon...
December 23, 2016: Nature Communications
https://www.readbyqxmd.com/read/27995908/structural-insights-into-ribosomal-rescue-by-dom34-and-hbs1-at-near-atomic-resolution
#11
Tarek Hilal, Hiroshi Yamamoto, Justus Loerke, Jörg Bürger, Thorsten Mielke, Christian M T Spahn
The surveillance of mRNA translation is imperative for homeostasis. Monitoring the integrity of the message is essential, as the translation of aberrant mRNAs leads to stalling of the translational machinery. During ribosomal rescue, arrested ribosomes are specifically recognized by the conserved eukaryotic proteins Dom34 and Hbs1, to initiate their recycling. Here we solve the structure of Dom34 and Hbs1 bound to a yeast ribosome programmed with a nonstop mRNA at 3.3 Å resolution using cryo-electron microscopy...
December 20, 2016: Nature Communications
https://www.readbyqxmd.com/read/27986852/structure-of-a-30s-pre-initiation-complex-stalled-by-ge81112-reveals-structural-parallels-in-bacterial-and-eukaryotic-protein-synthesis-initiation-pathways
#12
Jorge P López-Alonso, Attilio Fabbretti, Tatsuya Kaminishi, Idoia Iturrioz, Letizia Brandi, David Gil-Carton, Claudio O Gualerzi, Paola Fucini, Sean R Connell
In bacteria, the start site and the reading frame of the messenger RNA are selected by the small ribosomal subunit (30S) when the start codon, typically an AUG, is decoded in the P-site by the initiator tRNA in a process guided and controlled by three initiation factors. This process can be efficiently inhibited by GE81112, a natural tetrapeptide antibiotic that is highly specific toward bacteria. Here GE81112 was used to stabilize the 30S pre-initiation complex and obtain its structure by cryo-electron microscopy...
December 15, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27980192/rescuing-stalled-ribosomes
#13
EDITORIAL
Valda Vinson
No abstract text is available yet for this article.
December 16, 2016: Science
https://www.readbyqxmd.com/read/27934701/translational-termination-without-a-stop-codon
#14
Nathan R James, Alan Brown, Yuliya Gordiyenko, V Ramakrishnan
Ribosomes stall when they encounter the end of messenger RNA (mRNA) without an in-frame stop codon. In bacteria, these "nonstop" complexes can be rescued by alternative ribosome-rescue factor A (ArfA). We used electron cryomicroscopy to determine structures of ArfA bound to the ribosome with 3'-truncated mRNA, at resolutions ranging from 3.0 to 3.4 angstroms. ArfA binds within the ribosomal mRNA channel and substitutes for the absent stop codon in the A site by specifically recruiting release factor 2 (RF2), initially in a compact preaccommodated state...
December 16, 2016: Science
https://www.readbyqxmd.com/read/27920294/effects-of-increasing-the-affinity-of-card-for-rna-polymerase-on-mycobacterium-tuberculosis-growth-rrna-transcription-and-virulence
#15
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 that stimulates formation of RNAP-promoter open complexes. CarD plays a complex role in M. tuberculosis growth and virulence that is not fully understood. Therefore, to gain further insight into the role of CarD in M. tuberculosis 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...
February 15, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/27906650/structure-of-the-70s-ribosome-from-human-pathogen-staphylococcus-aureus
#16
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
#17
Paul Huter, Claudia Müller, Bertrand Beckert, Stefan Arenz, Otto Berninghausen, Roland Beckmann, Daniel N Wilson
In bacteria, ribosomes stalled on truncated mRNAs that lack a stop codon are rescued by the transfer-messenger RNA (tmRNA), alternative rescue factor A (ArfA) or ArfB systems. Although tmRNA-ribosome and ArfB-ribosome structures have been determined, how ArfA recognizes the presence of truncated mRNAs and recruits the canonical termination release factor RF2 to rescue the stalled ribosomes is unclear. Here we present a cryo-electron microscopy reconstruction of the Escherichia coli 70S ribosome stalled on a truncated mRNA in the presence of ArfA and RF2...
January 26, 2017: Nature
https://www.readbyqxmd.com/read/27906160/mechanistic-insights-into-the-alternative-translation-termination-by-arfa-and-rf2
#18
Chengying Ma, Daisuke Kurita, Ningning Li, Yan Chen, Hyouta Himeno, Ning Gao
During cellular translation of messenger RNAs by ribosomes, the translation apparatus sometimes pauses or stalls at the elongation and termination steps. With the exception of programmed stalling, which is usually used by cells for regulatory purposes, ribosomes stalled on mRNAs need to be terminated and recycled to maintain adequate translation capacity. Much ribosome stalling originates in aberrant mRNAs that lack a stop codon. Transcriptional errors, misprocessing of primary transcripts, and undesired mRNA cleavage all contribute to the formation of non-stop mRNAs...
January 26, 2017: Nature
https://www.readbyqxmd.com/read/27892503/trans-translation-is-essential-in-the-human-pathogen-legionella-pneumophila
#19
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
#20
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
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