scholarly journals The Archaeal Elongation Factor EF-2 Induces the Release of aIF6 From 50S Ribosomal Subunit

2021 ◽  
Vol 12 ◽  
Author(s):  
Giada Lo Gullo ◽  
Maria Luisa De Santis ◽  
Alessandro Paiardini ◽  
Serena Rosignoli ◽  
Alice Romagnoli ◽  
...  
Keyword(s):  
Large Subunit ◽  
Ribosomal Subunit ◽  
Elongation Factor ◽  
Structural Data ◽  
Small Subunit ◽  
Large Ribosomal Subunit ◽  
Homologous Proteins ◽  
Gtp Hydrolysis ◽  
Elongation Factor 2 ◽  
Docking Protein

The translation factor IF6 is a protein of about 25 kDa shared by the Archaea and the Eukarya but absent in Bacteria. It acts as a ribosome anti-association factor that binds to the large subunit preventing the joining to the small subunit. It must be released from the large ribosomal subunit to permit its entry to the translation cycle. In Eukarya, this process occurs by the coordinated action of the GTPase Efl1 and the docking protein SBDS. Archaea do not possess a homolog of the former factor while they have a homolog of SBDS. In the past, we have determined the function and ribosomal localization of the archaeal (Sulfolobus solfataricus) IF6 homolog (aIF6) highlighting its similarity to the eukaryotic counterpart. Here, we analyzed the mechanism of aIF6 release from the large ribosomal subunit. We found that, similarly to the Eukarya, the detachment of aIF6 from the 50S subunit requires a GTPase activity which involves the archaeal elongation factor 2 (aEF-2). However, the release of aIF6 from the 50S subunits does not require the archaeal homolog of SBDS, being on the contrary inhibited by its presence. Molecular modeling, using published structural data of closely related homologous proteins, elucidated the mechanistic interplay between the aIF6, aSBDS, and aEF2 on the ribosome surface. The results suggest that a conformational rearrangement of aEF2, upon GTP hydrolysis, promotes aIF6 ejection. On the other hand, aSBDS and aEF2 share the same binding site, whose occupation by SBDS prevents aEF2 binding, thereby inhibiting aIF6 release.

A Possible Role for 5 S rRNA as a Bridge Between Ribosomal Subunits

1973 ◽  
Vol 51 (12) ◽  
pp. 1669-1672 ◽  
Author(s):  
A. A. Azad ◽  
B. G. Lane
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Large Subunit ◽  
Ribosomal Subunit ◽  
Small Subunit ◽  
Low Molecular Weight ◽  
Large Ribosomal Subunit ◽  
Ribosomal Subunits ◽  
Wheat Embryo ◽  
Present Context

18 S rRNA is a high molecular weight polyribonucleotide found in the small subunit, and 26 S rRNA is a high molecular weight polyribonucleotide found in the large subunit, whereas 5 S rRNA and 5.8 S rRNA are low molecular weight ("satellite") polyribonucleotides confined to the large subunit of wheat-embryo ribosomes. Under the same conditions in which 5.8 S rRNA is known to complex efficiently and preferentially with 26 S rRNA, it has been observed that 5 S rRNA complexes efficiently and preferentially with 18 S rRNA. Since 5 S rRNA is a component of the large ribosomal subunit, but it complexes preferentially with 18 S rRNA, which is a component of the small ribosomal subunit, it has been proposed that 5 S rRNA may serve as a "bridge" to mediate reversible association between the small and large ribosomal subunits. The possible role that a polycistronic precursor of rRNA might be visualized to play in the biogenesis and assembly of reversibly associating ribosomal subunits is alluded to in the present context.

scholarly journals Characterization of the nuclear export adaptor protein Nmd3 in association with the 60S ribosomal subunit

2010 ◽  
Vol 189 (7) ◽  
pp. 1079-1086 ◽  
Author(s):  
Jayati Sengupta ◽  
Cyril Bussiere ◽  
Jesper Pallesen ◽  
Matthew West ◽  
Arlen W. Johnson ◽  
...  
Keyword(s):  
Binding Site ◽  
Nuclear Export ◽  
Large Subunit ◽  
Ribosomal Subunit ◽  
Adaptor Protein ◽  
Structural Data ◽  
Release Mechanism ◽  
Nuclear Pore ◽  
Subunit Joining

The nucleocytoplasmic shuttling protein Nmd3 is an adaptor for export of the 60S ribosomal subunit from the nucleus. Nmd3 binds to nascent 60S subunits in the nucleus and recruits the export receptor Crm1 to facilitate passage through the nuclear pore complex. In this study, we present a cryoelectron microscopy (cryo-EM) reconstruction of the 60S subunit in complex with Nmd3 from Saccharomyces cerevisiae. The density corresponding to Nmd3 is directly visible in the cryo-EM map and is attached to the regions around helices 38, 69, and 95 of the 25S ribosomal RNA (rRNA), the helix 95 region being adjacent to the protein Rpl10. We identify the intersubunit side of the large subunit as the binding site for Nmd3. rRNA protection experiments corroborate the structural data. Furthermore, Nmd3 binding to 60S subunits is blocked in 80S ribosomes, which is consistent with the assigned binding site on the subunit joining face. This cryo-EM map is a first step toward a molecular understanding of the functional role and release mechanism of Nmd3.

The ribosome as a molecular machine: the mechanism of tRNA–mRNA movement in translocation

2011 ◽  
Vol 39 (2) ◽  
pp. 658-662 ◽  
Author(s):  
Marina V. Rodnina ◽  
Wolfgang Wintermeyer
Keyword(s):  
Ribosomal Subunit ◽  
Elongation Factor ◽  
Thermal Fluctuations ◽  
Spontaneous Movement ◽  
Conformational States ◽  
Gtp Hydrolysis ◽  
Time Resolved ◽  
Directional Bias ◽  
Loosely Coupled ◽  
Dynamic Events

Translocation of tRNA and mRNA through the ribosome is one of the most dynamic events during protein synthesis. In the cell, translocation is catalysed by EF-G (elongation factor G) and driven by GTP hydrolysis. Major unresolved questions are: how the movement is induced and what the moving parts of the ribosome are. Recent progress in time-resolved cryoelectron microscopy revealed trajectories of tRNA movement through the ribosome. Driven by thermal fluctuations, the ribosome spontaneously samples a large number of conformational states. The spontaneous movement of tRNAs through the ribosome is loosely coupled to the motions within the ribosome. EF-G stabilizes conformational states prone to translocation and promotes a conformational rearrangement of the ribosome (unlocking) that accelerates the rate-limiting step of translocation: the movement of the tRNA anticodons on the small ribosomal subunit. EF-G acts as a Brownian ratchet providing directional bias for movement at the cost of GTP hydrolysis.

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 Structural consequences of the interaction of RbgA with a 50S ribosomal subunit assembly intermediate

2019 ◽  
Vol 47 (19) ◽  
pp. 10414-10425 ◽  
Author(s):  
Amal Seffouh ◽  
Nikhil Jain ◽  
Dushyant Jahagirdar ◽  
Kaustuv Basu ◽  
Aida Razi ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Large Subunit ◽  
Ribosomal Subunit ◽  
Catalytic Residue ◽  
Gtp Hydrolysis ◽  
Subunit Assembly ◽  
Cryo Electron Microscopy ◽  
Assembly Factor ◽  
Assembly Intermediate ◽  
Immature Particle

Abstract Bacteria harbor a number GTPases that function in the assembly of the ribosome and are essential for growth. RbgA is one of these GTPases and is required for the assembly of the 50S subunit in most bacteria. Homologs of this protein are also implicated in the assembly of the large subunit of the mitochondrial and eukaryotic ribosome. We present here the cryo-electron microscopy structure of RbgA bound to a Bacillus subtilis 50S subunit assembly intermediate (45SRbgA particle) that accumulates in cells upon RbgA depletion. Binding of RbgA at the P site of the immature particle stabilizes functionally important rRNA helices in the A and P-sites, prior to the completion of the maturation process of the subunit. The structure also reveals the location of the highly conserved N-terminal end of RbgA containing the catalytic residue Histidine 9. The derived model supports a mechanism of GTP hydrolysis, and it shows that upon interaction of RbgA with the 45SRbgA particle, Histidine 9 positions itself near the nucleotide potentially acting as the catalytic residue with minimal rearrangements. This structure represents the first visualization of the conformational changes induced by an assembly factor in a bacterial subunit intermediate.

Binding of GDP to a ribosomal protein after elongation factor-2 dependent GTP hydrolysis

1992 ◽  
Vol 1132 (3) ◽  
pp. 284-289 ◽  
Author(s):  
Jean-Pierre Lavergne ◽  
Anne-Marie Reboud ◽  
Bruno Sontag ◽  
Dominique Guillot ◽  
Jean-Paul Reboud
Keyword(s):  
Ribosomal Protein ◽  
Elongation Factor ◽  
Gtp Hydrolysis ◽  
Elongation Factor 2

scholarly journals Five Fungal Pathogens Are Responsible for Bayberry Twig Blight and Fungicides Were Screened for Disease Control

2020 ◽  
Vol 8 (5) ◽  
pp. 689 ◽  
Author(s):  
Wenjun Li ◽  
Ming Hu ◽  
Yang Xue ◽  
Zhijun Li ◽  
Yanfei Zhang ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Fungal Pathogens ◽  
Large Subunit ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Its Region ◽  
Small Subunit ◽  
Field Application ◽  
Financial Loss ◽  
Twig Blight

Bayberry (Myrica rubra) is a commercial fruit in China. For the past seven years, twig blight disease has been attacking bayberry plantations in Shantou City, Guangdong Province, China, leading to destructive damage and financial loss. In this study, five fungal species associated with twig dieback and stem blight were identified based on morphological characteristics combined with multilocus sequence analysis (MLSA) on the internal transcribed spacer (ITS) region, partial sequences of β-tubulin (tub2), translation elongation factor 1-α (tef1-α), large subunit ribosomal RNA (LSU) and small subunit ribosomal RNA (SSU) genes, which are Epicoccum sorghinum, Neofusicoccum parvum, Lasiodiplodia theobromae, Nigrospora oryzae and a Pestalotiopsis new species P. myricae. P. myricae is the chief pathogen in fields, based on its high isolation rate and fast disease progression after inoculation. To our knowledge, this is the first study reporting the above five fungi as the pathogens responsible for bayberry twig blight. Indoor screening of fungicides indicates that Prochloraz (copper salt) is the most promising fungicide for field application, followed by Pyraclostrobin, 15% Difenoconazole + 15% Propiconazole, Difenoconazole and Myclobutanil. Additionally, Bacillus velezensis strain 3–10 and zeamines from Dickeya zeae strain EC1 could be used as potential ecofriendly alternatives to control the disease.

Purification of the large ribosomal subunit via its association with the small subunit

2009 ◽  
Vol 395 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Samuel P. Simons ◽  
Thomas J. McLellan ◽  
Paul A. Aeed ◽  
Richard P. Zaniewski ◽  
Charlene R. Desbonnet ◽  
...  
Keyword(s):  
Ribosomal Subunit ◽  
Small Subunit ◽  
Large Ribosomal Subunit

scholarly journals Initiation factor 2 stabilizes the ribosome in a semirotated conformation

2015 ◽  
Vol 112 (52) ◽  
pp. 15874-15879 ◽  
Author(s):  
Clarence Ling ◽  
Dmitri N. Ermolenko
Keyword(s):  
Single Molecule ◽  
Translational Control ◽  
Large Subunit ◽  
Ribosomal Subunit ◽  
Bound State ◽  
Initiation Factor ◽  
Large Ribosomal Subunit ◽  
Control Of Gene Expression ◽  
Initiation Phase ◽  
Intersubunit Rotation

Intersubunit rotation and movement of the L1 stalk, a mobile domain of the large ribosomal subunit, have been shown to accompany the elongation cycle of translation. The initiation phase of protein synthesis is crucial for translational control of gene expression; however, in contrast to elongation, little is known about the conformational rearrangements of the ribosome during initiation. Bacterial initiation factors (IFs) 1, 2, and 3 mediate the binding of initiator tRNA and mRNA to the small ribosomal subunit to form the initiation complex, which subsequently associates with the large subunit by a poorly understood mechanism. Here, we use single-molecule FRET to monitor intersubunit rotation and the inward/outward movement of the L1 stalk of the large ribosomal subunit during the subunit-joining step of translation initiation. We show that, on subunit association, the ribosome adopts a distinct conformation in which the ribosomal subunits are in a semirotated orientation and the L1 stalk is positioned in a half-closed state. The formation of the semirotated intermediate requires the presence of an aminoacylated initiator, fMet-tRNAfMet, and IF2 in the GTP-bound state. GTP hydrolysis by IF2 induces opening of the L1 stalk and the transition to the nonrotated conformation of the ribosome. Our results suggest that positioning subunits in a semirotated orientation facilitates subunit association and support a model in which L1 stalk movement is coupled to intersubunit rotation and/or IF2 binding.

Sign in / Sign up

Export Citation Format

Share Document