Stress-stiffening-mediated stem-cell commitment switch in soft responsive hydrogels.

Bulk matrix stiffness has emerged as a key mechanical cue in stem cell differentiation. Here, we show that the commitment and differentiation of human mesenchymal stem cells encapsulated in physiologically soft (∼0.2-0.

Direct observation of the translocation mechanism of transcription termination factor Rho.

Rho is a ring-shaped, ATP-fueled motor essential for remodeling transcriptional complexes and R-loops in bacteria. Despite years of research on this fundamental model helicase, key aspects of its mechanism of translocation remain largely unknown. Here, we used single-molecule manipulation and fluorescence methods to directly monitor the dynamics of RNA translocation by Rho.

Constructing a magnetic tweezers to monitor RNA translocation at the single-molecule level.

Single-molecule methods have become an invaluable tool in the investigation of the mechanisms of nucleic-acid motors. Magnetic tweezers is a single-molecule manipulation technique that permits the real-time measurement of enzyme activities on single nucleic-acid molecules at high-resolution, high-throughput, and inherently constant force. Here, we describe several aspects of the implementation of magnetic tweezers, with special emphasis on the construction of a simple magnetic trap and, in particular, on the detailed description of image analysis methods to measure the extension changes in nucleic-acid molecules induced by protein activity.

Membrane nanowaves in single and collective cell migration.

We report the characterization of three-dimensional membrane waves for migrating single and collective cells and describe their propagation using wide-field optical profiling technique with nanometer resolution. We reveal the existence of small and large membrane waves the amplitudes of which are in the range of ∼ 3-7 nm to ∼ 16-25 nm respectively, through the cell. For migrating single-cells, the amplitude of these waves is about 30 nm near the cell edge.

Mutagenesis-based evidence for an asymmetric configuration of the ring-shaped transcription termination factor Rho.

Transcription termination factor Rho is an ATP-dependent ring-shaped molecular motor that tracks along RNA to dissociate RNA-DNA duplexes and transcription complexes in its path. The Rho hexamer contains two distinct sites for interaction with RNA. The primary binding site is composed of pyrimidine-specific binding clefts that are located in the N-terminal domains and anchor Rho to transcripts at C-rich Rut (Rho utilization) sites.