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Translocation of the tRNA·mRNA complex through the bacterial ribosome is driven by the multidomain guanosine triphosphatase elongation factor G (EF-G). We have used isothermal titration calorimetry to characterize the binding of GDP and GTP to free EF-G at 4°C, 20°C, and 37°C. The binding affinity of EF-G is higher to GDP than to GTP at 4°C, but lower at 37°C. The binding enthalpy and entropy chan
During the stringent response, Escherichia coli enzyme RelA produces the ppGpp alarmone, which in turn regulates transcription, translation and replication. We show that ppGpp dramatically increases the turnover rate of its own ribosome-dependent synthesis by RelA, resulting in direct positive regulation of an enzyme by its product. Positive allosteric regulation therefore constitutes a new mechan
RelE/RelB is a well-characterized toxin-anti-toxin pair involved in nutritional stress responses in Bacteria and Archae. RelE lacks any eukaryote homolog, but we demonstrate here that it efficiently and specifically cleaves mRNA in the A site of the eukaryote ribosome. The cleavage mechanism is similar to that in bacteria, showing the feasibility of A-site cleavage of mRNA for regulatory purposes
In a recent issue of Molecular Cell, Studer and Joseph (2006) clarified the molecular basis of the "standby model" and demonstrated how two-step binding of mRNAs to the small ribosomal subunit depends on initiator tRNA and initiation factors, with far-reaching implications for posttranscriptional regulation of gene expression.
This theoretical work covers structural and biochemical aspects of nucleotide binding and GDP/GTP exchange of GTP hydrolases belonging to the family of small GTPases. Current models of GDP/GTP exchange regulation are often based on two specific assumptions. The first is that the conformation of a GTPase is switched by the exchange of the bound nucleotide from GDP to GTP or vice versa. The second i
Bacterial protein synthesis involves four protein factors that belong to the GTPase family: IF2, EF-G, EF-Tu, and RF3. Their role in translation has attracted considerable interest over the recent decades. Cryoelectron microscopy has made it possible to monitor the dynamics of the ribosome upon binding of the translation factors, and biochemical findings have associated the structural data with fu
In eukaryotes, termination of mRNA translation is triggered by the essential polypeptide chain release factors eRF1, recognizing all three stop codons, and eRF3, a member of the GTPase superfamily with a role that has remained opaque. We have studied the kinetic and thermodynamic parameters of the interactions between eRF3 and GTP, GDP and the non-hydrolysable GTP analogue GDPNP in the presence (K
Background. During the translation of mRNA into polypeptide, elongation factor G (EF-G) catalyzes the translocation of peptidyl-tRNA from the A site to the P site of the ribosome. According to the 'classical' model, EF-G in the GTP-bound form promotes translocation, while hydrolysis of the bound GTP promotes dissociation of the factor from the post-translocation ribosome. According to a more recen
After peptide release by a class-1 release factor, the ribosomal subunits must be recycled back to initiation. We have demonstrated that the distance between a strong Shine-Dalgarno (SD) sequence and a codon in the P site is crucial for the binding stability of the deacylated tRNA in the P site of the posttermination ribosome and the in-frame maintenance of its mRNA. We show that the elongation fa
RNA polymerases differ functionally and structurally in the initiation phase of transcription, when polymerization of 8-12 nucleotides occurs, from the later phases of transcription. Here we argue that the ribosome also might have different properties when translating the first codons in open reading frames, as compared with the later phases of translation.
#108. Entrelacé, for four vibraphonists on one vibraphone
Mesenchymal stem cells (MSCs) were proposed as a critical therapeutic candidate in diabetic nephropathy (DN). Renal stem cells as a source for repairing are controversial. The purpose of the present study was to evaluate the effect of kidney rat stem cells on DN. After evaluation of surface stem cell markers by flow cytometry analysis, MSCs that were labeled with CM-Dil were injected to experiment
Introduction: Mesenchymal stem cells (MSCs) were proposed as a critical therapeutic candidate in diabetic nephropathy (DN). Renal stem cells as a source for repairing are controversial. The purpose of the present study was to evaluate the effect of kidney rat stem cells on DN. Materials and methods: After separation of renal stem cells from rat kidney, the surface Stem cell markers were assessed b
This conceptual paper addresses how the emergent impact agenda is slowly but surely changing the normative framework of modern science. In order to understand such a cultural shift, we draw a chronology around the evaluation regime of research impact and identify a causal mechanism that changes the disciplinary structure of the research ecosystem. We draw upon a sociological model of scientific kn
