A5019332732- RNA Interference and Gene Delivery
- RNA and protein synthesis mechanisms
- RNA Research and Splicing
- Bacteriophages and microbial interactions
- Advanced biosensing and bioanalysis techniques
- RNA modifications and cancer
- Bacterial Genetics and Biotechnology
- CRISPR and Genetic Engineering
- Cancer Cells and Metastasis
- Pluripotent Stem Cells Research
- Glioma Diagnosis and Treatment
- Advanced Biosensing Techniques and Applications
- Enzyme Production and Characterization
- Advanced Fluorescence Microscopy Techniques
- DNA Repair Mechanisms
- Click Chemistry and Applications
- Virus-based gene therapy research
- Chemical Synthesis and Analysis
- DNA and Nucleic Acid Chemistry
- Mitochondrial Function and Pathology
University of Regensburg
2016-2023
University Hospital Schleswig-Holstein
2013-2016
University of Lübeck
2010-2016
Significance Argonaute (Ago) proteins are the key component of RNA-induced silencing complex (RISC) and mediate RNA interference (RNAi) in association with small RNAs. Although overall mechanism RNAi is well understood, many molecular details this process not. The implementation pre-steady-state kinetic techniques enabled us to develop a comprehensive minimal mechanistic model for human Ago 2 (hAgo2)-mediated target binding cleavage, dissecting binary as ternary formation cleavage into...
Abstract Human Argonaute 2 (hAgo2) constitutes the functional core of RNA interference pathway. Guide RNAs direct hAgo2 to target mRNAs, which ultimately leads hAgo2-mediated mRNA degradation or translational inhibition. Here, we combine site-specifically labeled with time-resolved single-molecule FRET measurements monitor conformational states and dynamics hAgo2-RNA complexes in solution that remained elusive so far. We observe dynamic anchoring release guide’s 3’-end from PAZ domain during...
Argonaute (Ago) proteins from all three domains of life are key players in processes that specifically regulate cellular nucleic acid levels. Some these Ago proteins, among them human Argonaute2 (hAgo2) and the archaeal organism Methanocaldococcus jannaschii (MjAgo), able to cleave target strands recognised via an Ago-associated complementary guide strand. Here we present in-depth kinetic side-by-side analysis hAgo2 MjAgo substrate binding as well strand cleavage, which enabled us disclose...
The human Argonaute 2 (hAgo2) protein is a key player of RNA interference (RNAi). Upon complex formation with small non-coding RNAs, the initially interacts 5′-end given guide through multiple interactions within MID domain. This interaction has been reported to show strong bias for U and A over C G at 5′-position. Performing molecular dynamics simulations binary hAgo2/OH–guide–RNA complexes, we that hAgo2 highly flexible capable binding strands all four possible 5′-bases. Especially, in...
Abstract Human cells are complex entities in which molecular recognition and selection critical for cellular processes often driven by structural changes dynamic interactions. Biomolecules appear different chemical states, modifications, such as phosphorylation, affect their function. Hence, using proteins chemically native state biochemical biophysical assays is essential. Single‐molecule FRET measurements allow exploration of the structure, function dynamics biomolecules but cannot be...
The discovery of RNA interference (RNAi) gave rise to the development new nucleic acid-based technologies as powerful investigational tools and potential therapeutics. Mechanistic key details RNAi in humans need be deciphered yet, before such approaches take root biomedicine molecular therapy.
Sm-like (Lsm) proteins are found in all three domains of life. They crucially involved the RNA metabolism prokaryotic organisms. To exert their function, they assemble into hexa- or heptameric rings and bind via a conserved binding pocket for uridine stretches inner pore ring. Despite secondary structure Lsm proteins, there several features that lead to structural diversification this protein family mediates participation variety processes related metabolism. Until recently, cellular...
Posttranscriptional processes in Bacteria include the association of small regulatory RNAs (sRNA) with a target mRNA. The sRNA/mRNA annealing process is often mediated by an RNA chaperone called Hfq. functional role bacterial and eukaryotic Lsm proteins partially understood, whereas knowledge about archaeal scarce. Here, we used genetically tractable hyperthermophile Pyrococcus furiosus to identify protein interaction partners Sm-like (PfuSmAP1) using mass spectrometry performed...
The human TAR RNA-binding protein (hTRBP) and activator of kinase R (hPACT) are important players in RNA interference (RNAi). Together with hArgonaute2 (hAgo2) hDicer they have been reported to form the RISC-loading complex (RLC). Among other functions, hTRBP was suggested assist loading hAgo2 small interfering RNAs (siRNAs) within RLC. Although several studies conducted evaluate specific functions hPACT RNAi, exact mechanisms modes action still unknown. Here, we present a biochemical study...
Abstract Cellular nucleic acid levels are specifically regulated by Argonaute (Ago) proteins. Eukaryotic (e)Ago participates mainly in posttranscriptional regulation of gene expression. In contrast, prokaryotic (p)Ago proteins involved defense processes. Our work adds to the understanding pAgo function and elucidates dynamic aspects Ago-mediated substrate binding thereby highlighting mechanistic differences between eAgo