Abstract Assembly of 30S ribosomes involves the hierarchical addition ribosomal proteins that progressively stabilize folded 16S rRNA. Here, we use three-color single molecule FRET to show how combinations uS4, uS17 and bS20 in 5′ domain enable recruitment protein bS16, next join complex. Analysis real-time bS16 binding events shows binds both native non-native forms The rRNA conformation is increasingly favored after binds, explaining drives later steps assembly. Chemical footprinting...

The (1)H NMR spectra of RNAs representing E. coli 23S rRNA helix 69 with [1,3-(15)N]pseudouridine modification at specific sites reveal unique roles for pseudouridine in stabilizing base-stacking interactions the hairpin loop region.

Assembly of the bacterial ribosomal small subunit (SSU) begins with folding five-way junction upon interaction primary binding protein S4. This complex contains largest contiguous molecular signature, which is a conserved feature in all 16S rRNAs. In previous study, we used all-atom dynamics simulations to demonstrate that co-evolving signature N-terminus S4 intrinsically disordered and capable accelerating process through fly casting mechanism. this paper, comparisons between MD FRET...

Helix 69 in 23S rRNA is a region the ribosome that participates considerable number of RNA–RNA and RNA–protein interactions. Conformational flexibility essential for such to interact accommodate protein factors at different stages biosynthesis. In this study, pH-dependent structural stability changes were observed helix through variety spectroscopic techniques, as circular dichroism spectroscopy, UV melting, nuclear magnetic resonance spectroscopy. Escherichia coli rRNA, contains...

Loop conformation: The loop of the RNA domain helix 69 (H69) was modified with fluorescent analogue 2-aminopurine (2AP), thus showing different conformational states under various conditions. application this model reveals unique impact aminoglycoside neomycin, which differs from effects structurally related compounds paromomycin and gentamicin, on H69 conformation in solution (see picture).

Ribosomal protein S4 nucleates assembly of the 30S ribosome 5′ and central domains, which is crucial for survival cells. Protein changes structure its 16S rRNA binding site, passing through a non-native intermediate complex before forming native S4-rRNA contacts. Ensemble FRET was used to measure thermodynamic stability complexes in presence Mg 2+ ions other 5′-domain proteins. Equilibrium titrations Cy3-labeled RNA with Cy5-labeled showed that preferentially stabilize complex. In contrast,...

Single-stranded telomeric overhangs are ∼200 nucleotides long and can form tandem G-quadruplex (GQ) structures, which reduce their accessibility to nucleases proteins that activate DNA damage response. Whether these GQs further stack compact superstructures, may provide better protection for longer telomeres, is not known. We report single-molecule measurements where the of 24-144 nucleotide human molecules interrogated by a short PNA molecule complementary single GGGTTA repeat, as...

Ribosomes are ribonucleoprotein particles that essential for protein biosynthesis in all forms of life. During ribosome biogenesis, transcription, folding, modification, and processing rRNA coupled to the assembly proteins. Various factors required synchronize different processes occur during biogenesis. Herein, RNA chaperone strand annealing activity modification enzyme small subunit methyltransferase C (RsmC), which modifies guanine 2-methylguanosine (m2 G) at position 1207 16S...

Tertiary sequence motifs encode interactions between RNA helices that create the three-dimensional structures of ribosomal subunits. A Right Angle motif at junction 16S 5 and 6 (J5/6) is universally conserved amongst small subunit rRNAs forms a stable right angle in minimal RNAs. J5/6 does not form mature ribosome, suggesting this encodes metastable structure needed for ribosome biogenesis. In study, mutations block 30S assembly maturation Escherichia coli. Folding assays in-cell X-ray...

Abstract Glycogen synthase kinase 3 (GSK3) was identified as an enzyme regulating sperm protein phosphatase. The GSK3α paralog, but not GSK3β, is essential for function. Sperm lacking display altered motility and are unable to undergo hyperactivation, which fertilization. Male mice sperm‐specific calcineurin (PP2B), a calcium regulated phosphatase, in testis sperm, also infertile. Loss of PP2B results impaired epididymal maturation motility. phenotypes knockout similar, prompting us examine...

Despite the advances in modern medicine, antibiotic resistance is a persistent and growing threat to world. Thus, discovery development of novel antibiotics have become crucial combat multi-drug resistant pathogens. The goal our research discover small molecular peptide that can disrupt synthesis new ribosomes. Using phage display technique, we discovered 7-mer binds second strand 16S h34 RNA with dissociation constant low micromolar range. Binding alters structure inhibits binding ribosomal...

Bacterial ribosome small subunit rRNA (16S rRNA) contains 11 nucleotide modifications scattered throughout all its domains. The 16S pseudouridylation enzyme, RsuA, which modifies U516, is a survival protein essential for bacterial under stress conditions. A comparison of the growth curves wildtype and RsuA knock-out E. coli strains illustrates that renders advantage to bacteria streptomycin stress. RsuA-dependent was found be dependent on activity. In addition, role as trans-acting factor...

The ribosome is the ribonucleoprotein machine that carries out protein biosynthesis in all forms of life. Perfect synchronization between ribosomal RNA (rRNA) transcription, folding, post-transcriptional modification, maturation, and assembly r-proteins essential for rapid formation structurally functionally accurate ribosomes. Many nucleotide modification enzymes may function as factors oversee accuracy assembly. RsmG a methyltransferase enzyme responsible N7 methylation G527 16S rRNA. Here...

The ribosome is a large ribonucleoprotein complex that synthesizes protein in all living organisms. Ribosome biogenesis process requires synchronization of various cellular events, including ribosomal RNA (rRNA) transcription, assembly, and processing post-transcriptional modification rRNA. fine-tuned with assembly factors, possibly nucleotide enzymes. Ribosomal small subunit pseudouridine synthase A (RsuA) pseudouridylates U516 16S helix 18. Protein RsuA multi-domain contains the N-terminal...

Abstract N6‐methyladenosine (m 6 A) is the most abundant nucleotide modification observed in eukaryotic mRNA. Changes m A levels transcriptome are tightly correlated to expression of methyltransferases and demethylases. Abnormal demethylases various diseases health conditions such as cancer, male infertility, obesity. This research explores efficacy A‐modified RNA an anticancer drug target. We discovered a 12‐mer peptide that binds specifically using phage display experiments. Our...

We performed single molecule studies to investigate the impact of several prominent small molecules (the oxazole telomestatin derivative L2H2-6OTD, pyridostatin, and Phen-DC3) on intermolecular G-quadruplex (i-GQ) formation between two guanine-rich DNA strands that had 3-GGG repeats in one strand 1-GGG repeat other (3+1 GGG), or 2-GGG each (2+2 GGG). Such structures are not only physiologically significant but have recently found use various biotechnology applications, ranging from DNA-based...

Schleifenkonformation: Die Schleife der RNA-Domäne Helix-69 (H69) wurde mit dem fluoreszierenden Adenin-Analogon 2-Aminopurin (2AP) modifiziert, was die je nach den Bedingungen unterschiedlichen Konformationszustände sichtbar machte. Verwendung dieser Modell-RNA zeigt besonderen Einfluss des Aminoglycosids Neomycin auf Schleifenkonformation von H69, sich strukturell ähnlichen Verbindungen Paromomycin und Gentamicin unterscheidet (siehe Bild). Detailed facts of importance to specialist...