scholarly journals Switch of the interactions between the ribosomal stalk and EF1A in the GTP- and GDP-bound conformations

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Kei Maruyama ◽  
Hirotatsu Imai ◽  
Momoko Kawamura ◽  
Sonoko Ishino ◽  
Yoshizumi Ishino ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Large Subunit ◽  
Elongation Factor ◽  
Binding Mode ◽  
Translation Elongation ◽  
Gtp Hydrolysis ◽  
Before And After ◽  
Ribosomal Stalk ◽  
Biochemical Analyses ◽  
Similar Affinity

Abstract Translation elongation factor EF1A delivers aminoacyl-tRNA to the ribosome in a GTP-bound form, and is released from the ribosome in a GDP-bound form. This association/dissociation cycle proceeds efficiently via a marked conformational change in EF1A. EF1A function is dependent on the ribosomal “stalk” protein of the ribosomal large subunit, although the precise mechanism of action of the stalk on EF1A remains unclear. Here, we clarify the binding mode of archaeal stalk aP1 to GTP-bound aEF1A associated with aPelota. Intriguingly, the C-terminal domain (CTD) of aP1 binds to aEF1A•GTP with a similar affinity to aEF1A•GDP. We have also determined the crystal structure of the aP1-CTD•aEF1A•GTP•aPelota complex at 3.0 Å resolution. The structure shows that aP1-CTD binds to a space between domains 1 and 3 of aEF1A. Biochemical analyses show that this binding is crucial for protein synthesis. Comparison of the structures of aP1-CTD•aEF1A•GTP and aP1-CTD•aEF1A•GDP demonstrates that the binding mode of aP1 changes markedly upon a conformational switch between the GTP- and GDP-bound forms of aEF1A. Taking into account biochemical data, we infer that aP1 employs its structural flexibility to bind to aEF1A before and after GTP hydrolysis for efficient protein synthesis.

Cytoskeletal proteins associate with components of the ribosomal maturation and translation apparatus in Xenopus stage I oocytes

Zygote ◽  
2014 ◽  
Vol 23 (5) ◽  
pp. 669-682 ◽  
Author(s):  
Loredana Chierchia ◽  
Margherita Tussellino ◽  
Domenico Guarino ◽  
Rosa Carotenuto ◽  
Nadia DeMarco ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Elongation Factor ◽  
Cytoskeletal Proteins ◽  
Stage I ◽  
Initiation Factor ◽  
Perinuclear Region ◽  
Eukaryotic Initiation Factor ◽  
Translation Apparatus ◽  
Ribosomal Stalk

SummaryActin-based cytoskeleton (CSK) and microtubules may bind to RNAs and related molecules implicated in translation. However, many questions remain to be answered regarding the role of cytoskeletal components in supporting the proteins involved in steps in the maturation and translation processes. Here, we performed co-immunoprecipitation and immunofluorescence to examine the association between spectrins, keratins and tubulin and proteins involved in 60S ribosomal maturation and translation in Xenopus stage I oocytes, including ribosomal rpl10, eukaryotic initiation factor 6 (Eif6), thesaurins A/B, homologs of the eEF1α elongation factor, and P0, the ribosomal stalk protein. We found that rpl10 and eif6 cross-reacted with the actin-based CSK and with tubulin. rpl10 co-localizes with spectrin, particularly in the perinuclear region. eif6 is similarly localized. Given that upon ribosomal maturation, the insertion of rpl10 into the 60S subunit occurs simultaneously with the release of eif6, one can hypothesise that actin-based CSK and microtubules provide the necessary scaffold for the insertion/release of these two molecules and, subsequently, for eif6 transport and binding to the mature 60S subunit. P0 and thesaurins cross-reacted with only spectrin and cytokeratins. Thesaurins aggregated at the oocyte periphery, rendering this a territory favourable site for protein synthesis; the CSK may support the interaction between thesaurins and sites of the translating ribosome. Moreover, given that the assembly of the ribosome stalk, where P0 is located, to the 60S subunit is essential for the release of eif6, it can be hypothesised that the CSK can facilitate the binding of the stalk to the 60S.

Affinities of the Boletus chromapes group to Royoungia and the description of two new genera, Harrya and Australopilus

2012 ◽  
Vol 25 (6) ◽  
pp. 418 ◽  
Author(s):  
Roy E. Halling ◽  
Mitchell Nuhn ◽  
Todd Osmundson ◽  
Nigel Fechner ◽  
James M. Trappe ◽  
...  
Keyword(s):  
New Genus ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Sister Group ◽  
Molecular Data ◽  
Translation Elongation ◽  
Spore Deposit ◽  
Large Subunit Rdna ◽  
Translation Elongation Factor 1Α

Harrya is described as a new genus of Boletaceae to accommodate Boletus chromapes, a pink-capped bolete with a finely scabrous stipe adorned with pink scabers, a chrome yellow base and a reddish-brown spore deposit. Phylogenetic analyses of large-subunit rDNA and translation elongation factor 1α confirmed Harrya as a unique generic lineage with two species, one of which is newly described (H. atriceps). Some Chinese taxa were recently placed in a separate genus, Zangia, supported by both morphology and molecular data. Multiple accessions from Queensland, Australia, support the synonymy of at least three species in a separate Australian clade in the new genus, Australopilus. The truffle-like Royoungia is also supported as a separate lineage in this clade of boletes. Even though it lacks stipe characters, it possesses the deep, bright yellow to orange pigments in the peridium. Additional collections from Zambia and Thailand represent independent lineages of uncertain phylogenetic placement in the Chromapes complex, but sampling is insufficient for formal description of new species. Specimens from Java referable to Tylopilus pernanus appear to be a sister group of the Harrya lineage.

scholarly journals Insights Into the Pathologic Roles and Regulation of Eukaryotic Elongation Factor-2 Kinase

2021 ◽  
Vol 8 ◽  
Author(s):  
Darby J. Ballard ◽  
Hao-Yun Peng ◽  
Jugal Kishore Das ◽  
Anil Kumar ◽  
Liqing Wang ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Protein Kinase ◽  
Elongation Factor ◽  
Protein Translation ◽  
Negative Regulator ◽  
Translation Elongation ◽  
Elongation Factor 2 ◽  
Global Rate ◽  
Eukaryotic Elongation Factor 2 ◽  
Eukaryotic Elongation Factor

Eukaryotic Elongation Factor-2 Kinase (eEF2K) acts as a negative regulator of protein synthesis, translation, and cell growth. As a structurally unique member of the alpha-kinase family, eEF2K is essential to cell survival under stressful conditions, as it contributes to both cell viability and proliferation. Known as the modulator of the global rate of protein translation, eEF2K inhibits eEF2 (eukaryotic Elongation Factor 2) and decreases translation elongation when active. eEF2K is regulated by various mechanisms, including phosphorylation through residues and autophosphorylation. Specifically, this protein kinase is downregulated through the phosphorylation of multiple sites via mTOR signaling and upregulated via the AMPK pathway. eEF2K plays important roles in numerous biological systems, including neurology, cardiology, myology, and immunology. This review provides further insights into the current roles of eEF2K and its potential to be explored as a therapeutic target for drug development.

Spegazzinia camelliae sp. nov. (Didymosphaeriaceae, Pleosprales), a new endophytic fungus from northern Thailand

Phytotaxa ◽  
2021 ◽  
Vol 483 (2) ◽  
pp. 117-128
Author(s):  
NAKARIN SUWANNARACH ◽  
JATURONG KUMLA ◽  
SAISAMORN LUMYONG
Keyword(s):  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Small Subunit ◽  
Internal Transcribed Spacers ◽  
Nuclear Ribosomal Dna ◽  
Full Description ◽  
Translation Elongation ◽  
Spine Length ◽  
Elongation Factor 1 Alpha

A new endophytic ascomycete, described herein as Spegazzinia camelliae, was isolated from leaves of Camellia sinensis var. assamica collected from Nan Province, Thailand. This species is characterized by basauxic conidiophores and dark brown to blackish brown α and β conidia. It can be distinguished from previously described Spegazzinia species by the spine length of the α conidia and the size of the β conidia. Multi-gene phylogenetic analyses of the small subunit (SSU), large subunit (LSU) and internal transcribed spacers (ITS) of the nuclear ribosomal DNA (rDNA) and the translation elongation factor 1-alpha (tef1) genes also support S. camelliae is a distinct new species within Spegazzinia. A full description, color photographs, illustrations and a phylogenetic tree showing the position of S. camelliae are provided.

scholarly journals Translation elongation factor 2 depletion by siRNA in mouse liver leads to mTOR-independent translational upregulation of ribosomal protein genes

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Maxim V. Gerashchenko ◽  
Mikhail V. Nesterchuk ◽  
Elena M. Smekalova ◽  
Joao A. Paulo ◽  
Piotr S. Kowalski ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Mammalian Cells ◽  
Ribosomal Proteins ◽  
Mouse Liver ◽  
Negative Impact ◽  
Elongation Factor ◽  
Ribosome Profiling ◽  
Translation Elongation Factor ◽  
Translation Elongation ◽  
Elongation Factor 2

Abstract Due to breakthroughs in RNAi and genome editing methods in the past decade, it is now easier than ever to study fine details of protein synthesis in animal models. However, most of our understanding of translation comes from unicellular organisms and cultured mammalian cells. In this study, we demonstrate the feasibility of perturbing protein synthesis in a mouse liver by targeting translation elongation factor 2 (eEF2) with RNAi. We were able to achieve over 90% knockdown efficacy and maintain it for 2 weeks effectively slowing down the rate of translation elongation. As the total protein yield declined, both proteomics and ribosome profiling assays showed robust translational upregulation of ribosomal proteins relative to other proteins. Although all these genes bear the TOP regulatory motif, the branch of the mTOR pathway responsible for translation regulation was not activated. Paradoxically, coordinated translational upregulation of ribosomal proteins only occurred in the liver but not in murine cell culture. Thus, the upregulation of ribosomal transcripts likely occurred via passive mTOR-independent mechanisms. Impaired elongation sequesters ribosomes on mRNA and creates a shortage of free ribosomes. This leads to preferential translation of transcripts with high initiation rates such as ribosomal proteins. Furthermore, severe eEF2 shortage reduces the negative impact of positively charged amino acids frequent in ribosomal proteins on ribosome progression.

Morphological characters and phylogenetic analysis reveal a new species of Phellinus with hooked hymenial setae from Vietnam

Phytotaxa ◽  
2018 ◽  
Vol 356 (1) ◽  
pp. 91 ◽  
Author(s):  
LIN ZHU ◽  
XING JI ◽  
JING SI ◽  
BAO-KAI CUI
Keyword(s):  
Rna Polymerase Ii ◽  
Large Subunit ◽  
Elongation Factor ◽  
Morphological Characters ◽  
Molecular Data ◽  
Translation Elongation ◽  
The Status ◽  
Large Subunit Rdna ◽  
Elongation Factor 1 ◽  
A New Species

Phellinus vietnamensis sp. nov. is described from Vietnam based on morphological characters and molecular data. Morphologically, it is characterized by perennial, pileate basidiomata, a dimitic hyphal system, hooked hymenial setae, and colorless, broadly subglobose to ovoid, thick-walled basidiospores 5.5–6 × 4.8–5.2 μm. Phylogenetically, the status of Phellinus vietnamensis is strongly supported based on sequences of the nuclear internal transcribed spacer (ITS) regions, the translation elongation factor 1-α gene (EF1-α) nuclear large subunit rDNA (nrLSU) and the second largest subunits of RNA polymerase II (RPB2).

scholarly journals Bambusicolous Arthrinium Species in Guangdong Province, China

2020 ◽  
Vol 11 ◽  
Author(s):  
Indunil C. Senanayake ◽  
Jayarama D. Bhat ◽  
Ratchadawan Cheewangkoon ◽  
Ning Xie
Keyword(s):  
Large Subunit ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Guangdong Province ◽  
Nuclear Ribosomal Dna ◽  
Internal Transcribed Spacer Region ◽  
Translation Elongation ◽  
Bambusicolous Fungi ◽  
Elongation Factor 1 Alpha ◽  
Elongation Factor 1

A survey of bambusicolous fungi in Bijiashan Mountain Park, Shenzhen, Guangdong Province, China, revealed several Arthrinium-like taxa from dead sheaths, twigs, and clumps of Bambusa species. Phylogenetic relationships were investigated based on morphology and combined analyses of the internal transcribed spacer region (ITS), large subunit nuclear ribosomal DNA (LSU), beta tubulin (β-tubulin), and translation elongation factor 1-alpha (tef 1-α) gene sequences. Based on morphological characteristics and phylogenetic data, Arthrinium acutiapicum sp. nov. and Arthrinium pseudorasikravindrae sp. nov. are introduced herein with descriptions and illustrations. Additionally, two new locality records of Arthrinium bambusae and Arthrinium guizhouense are described and illustrated.

scholarly journals Multi-gene phylogeny and taxonomy of Amauroderma s. lat. (Ganodermataceae)

2020 ◽  
Vol 44 (1) ◽  
pp. 206-239 ◽  
Author(s):  
Y.-F. Sun ◽  
D.H. Costa-Rezende ◽  
J.-H. Xing ◽  
J.-L. Zhou ◽  
B. Zhang ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Morphological Examination ◽  
Translation Elongation ◽  
Multiple Loci ◽  
Phylogenetic Studies ◽  
Molecular Phylogenetic

Amauroderma s.lat. has been defined mainly by the morphological features of non-truncate and double-walled basidiospores with a distinctly ornamented endospore wall. In this work, taxonomic and phylogenetic studies on species of Amauroderma s.lat. are carried out by morphological examination together with ultrastructural observations, and molecular phylogenetic analyses of multiple loci including the internal transcribed spacer regions (ITS), the large subunit of nuclear ribosomal RNA gene (nLSU), the largest subunit of RNA polymerase II (RPB1) and the second largest subunit of RNA polymerase II (RPB2), the translation elongation factor 1-α gene (TEF) and the β-tubulin gene (TUB). The results demonstrate that species of Ganodermataceae formed ten clades. Species previously placed in Amauroderma s.lat. are divided into four clades: Amauroderma s.str., Foraminispora, Furtadoa and a new genus Sanguinoderma. The classification of Amauroderma s. lat. is thus revised, six new species are described and illustrated, and eight new combinations are proposed. SEM micrographs of basidiospores of Foraminispora and Sanguinoderma are provided, and the importance of SEM in delimitation of taxa in this study is briefly discussed. Keys to species of Amauroderma s.str., Foraminispora, Furtadoa, and Sanguinoderma are also provided.

A new species and a new combination in the genus Aureoboletus (Boletales, Boletaceae) from southern China

Phytotaxa ◽  
2015 ◽  
Vol 222 (2) ◽  
pp. 129 ◽  
Author(s):  
Nian-Kai Zeng ◽  
MING ZHANG ◽  
ZHI-QUN Liang
Keyword(s):  
Rna Polymerase Ii ◽  
Southern China ◽  
Large Subunit ◽  
Line Drawing ◽  
Elongation Factor ◽  
Molecular Data ◽  
New Combination ◽  
Translation Elongation ◽  
Microscopic Features ◽  
A New Species

Two lineages of Aureoboletus (Boletales, Boletaceae) from southern China were revealed by using molecular data based on combined dataset of the nuclear ribosomal large subunit RNA (nrLSU), the translation elongation factor apha-1 (tef1-a) and the largest subunit of RNA polymerase II (rpb1). One of them corresponds with the previous morphology-based taxon, viz. Boletellus longicollis, another one is different from those taxa described based on morphological features. And, thus, Auroboletus clavatus sp. nov. and A. longicollis comb. nov. were proposed. A detailed description, colour photos of fresh basidiomata, and a line-drawing of microscopic features of the two taxa were provided.

scholarly journals Inhibition of protein synthesis in vitro by crotins and ricin. Effect on the steps of peptide chain elongation

1976 ◽  
Vol 156 (1) ◽  
pp. 7-13 ◽  
Author(s):  
S Sperti ◽  
L Montanaro ◽  
A Mattioli ◽  
G Testoni ◽  
F Stirpe
Keyword(s):  
Protein Synthesis ◽  
Rat Liver ◽  
Elongation Factor ◽  
Artemia Salina ◽  
Chain Elongation ◽  
Gtp Hydrolysis ◽  
Elongation Factor 2 ◽  
The Individual ◽  
Liver Ribosomes

The effects of crotin I and crotin II on the partial reactions of polypeptide chain elongation were investigated and compared with the known effects of ricin. Crotin II was a more powerful inhibitor than crotin I, but no qualitative differences between the two crotins were found. Rat liver ribosomes, preincubated with crotins and washed through sucrose gradients, remained inactive in protein synthesis. Among the individual steps of elongation, the peptidyltransferase reaction was unaffected by crotins, but some of the reactions that involve the interaction of elongation factors 1 and 2 with ribosomes were modified. A strong inhibition of the binding of elongation factor 2 to ribosomes and a stimulation of the elongation factor2-dependent GTP hydrolysis were observed; this indicates the formation of a very unstable elongation factor 2-GDP-ribosome complex, which, however, allows a single round of translocation to take place in the presence ofelongation factor 2 and added GTP. The elongation factor 1-dependent GTP hydrolysis was inhibited by crotins, whereas the enzymic binding of aminoacyl-tRNA, to both rat liver and Artemia salina ribosomes, was scarcely affected. In a protein-synthesizing system the inhibition by crotins and by ricin leads to a block of the nascent peptides on the ribosomal aminoacyl-tRNA site, an effect consistent with inhibition at the level of translocation. The mechanism of action of crotins appears to be very similar to that of ricin.

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