CDK1-CCNB1 creates a spindle checkpoint-permissive state by enabling MPS1 kinetochore localization.

Spindle checkpoint signaling is initiated by recruitment of the kinase MPS1 to unattached kinetochores during mitosis. We show that CDK1-CCNB1 and a counteracting phosphatase PP2A-B55 regulate the engagement of human MPS1 with unattached kinetochores by controlling the phosphorylation status of S281 in the kinetochore-binding domain. This regulation is essential for checkpoint signaling, since MPS1 is not recruited to unattached kinetochores and fails to support the recruitment of other checkpoint proteins.

Interlinked bistable mechanisms generate robust mitotic transitions.

The transitions between phases of the cell cycle have evolved to be robust and switch-like, which ensures temporal separation of DNA replication, sister chromatid separation, and cell division. Mathematical models describing the biochemical interaction networks of cell cycle regulators attribute these properties to underlying bistable switches, which inherently generate robust, switch-like, and irreversible transitions between states. We have recently presented new mathematical models for two control systems that regulate crucial transitions in the cell cycle: mitotic entry and exit, and the mitotic checkpoint.

CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening.

Background: The bacterial CRISPR system is fast becoming the most popular genetic and epigenetic engineering tool due to its universal applicability and adaptability. The desire to deploy CRISPR-based methods in a large variety of species and contexts has created an urgent need for the development of easy, time- and cost-effective methods enabling large-scale screening approaches.

Results: Here we describe CORALINA (comprehensive gRNA library generation through controlled nuclease activity), a method for the generation of comprehensive gRNA libraries for CRISPR-based screens.

A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit.

PP2A-B55 is one of the major phosphatases regulating cell division. Despite its importance for temporal control during mitotic exit, how B55 substrates are recognized and differentially dephosphorylated is unclear. Using phosphoproteomics combined with kinetic modeling to extract B55-dependent rate constants, we have systematically identified B55 substrates and assigned their temporal order in mitotic exit.

Premature Sister Chromatid Separation Is Poorly Detected by the Spindle Assembly Checkpoint as a Result of System-Level Feedback.

Sister chromatid cohesion, mediated by the cohesin complex, is essential for faithful mitosis. Nevertheless, evidence suggests that the surveillance mechanism that governs mitotic fidelity, the spindle assembly checkpoint (SAC), is not robust enough to halt cell division when cohesion loss occurs prematurely. The mechanism behind this poor response is not properly understood.

Tuning the dynamic range and sensitivity of optical oxygen-sensors by employing differently substituted polystyrene-derivatives.

Ten different polystyrene-derivatives were tested with respect to their potential use as matrix materials for optical oxygen sensors in combination with the platinum(II) meso-tetra(4-fluorophenyl)tetrabenzoporphyrin as indicator dye. Either halogen atoms or bulky residues were introduced as substituents on the phenyl ring. A fine-tuning of the sensor sensitivity was achieved, without compromising solubility of the indicator in the matrix by providing a chemical environment very similar to polystyrene (PS), a standard matrix in optical oxygen sensors.

Layer-by-layer processing and optical properties of core/alloy nanostructures.

A novel hydrothermal layer-by-layer processing method for the fabrication of core/alloy nanoparticles with highly tunable surface plasmon resonance is described. For a model system of Au/Au(x)Ag(1-x), the processing temperature, alloy composition, and alloy thickness resulted in unique and tailorable plasmonic signatures. The discrete dipole approximation and selective alloy etching were used to correlate this optical response with the particle morphology and alloy phase ultrastructure.