Fishnet mesh of centrin-Sfi1 drives ultrafast calcium-activated contraction of the giant cell .

is a unicellular ciliate capable of contracting to a quarter of its body length in less than five milliseconds. When measured as fractional shortening, this is an order of magnitude faster than motion powered by actomyosin. Myonemes, which are protein networks found near the cortex of many protists, are believed to power contraction.

A unified model for the dynamics of ATP-independent ultrafast contraction.

In nature, several ciliated protists possess the remarkable ability to execute ultrafast motions using protein assemblies called myonemes, which contract in response to Ca ions. Existing theories, such as actomyosin contractility and macroscopic biomechanical latches, do not adequately describe these systems, necessitating development of models to understand their mechanisms. In this study, we image and quantitatively analyze the contractile kinematics observed in two ciliated protists ( sp.

Mitosis: Augmin-based bridges keep kinetochores in line.

A recent study highlights the indispensability of the augmin complex for the construction of mitotic spindle bridging fibers, which in turn support accurate chromosome attachment and segregation.

Laser ablation reveals the impact of Cdc15p on the stiffness of the contractile ring.

The mechanics that govern the constriction of the contractile ring remain poorly understood yet are critical to understanding the forces that drive cytokinesis. We used laser ablation in fission yeast cells to unravel these mechanics focusing on the role of Cdc15p as a putative anchoring protein. Our work shows that the severed constricting contractile ring recoils to a finite point leaving a gap that can heal if less than ∼1 µm.

Force by minus-end motors Dhc1 and Klp2 collapses the S. pombe spindle after laser ablation.

A microtubule-based machine called the mitotic spindle segregates chromosomes when eukaryotic cells divide. In the fission yeast Schizosaccharomyces pombe, which undergoes closed mitosis, the spindle forms a single bundle of microtubules inside the nucleus. During elongation, the spindle extends via antiparallel microtubule sliding by molecular motors.

K-fiber bundles in the mitotic spindle are mechanically reinforced by Kif15.

The mitotic spindle, a self-constructed microtubule-based machine, segregates chromosomes during cell division. In mammalian cells, microtubule bundles called kinetochore fibers (k-fibers) connect chromosomes to the spindle poles. Chromosome segregation thus depends on the mechanical integrity of k-fibers.

Cytoskeletal biophysics: Passive crosslinker adapts to keep microtubule bundles on track.

Assembly of the mitotic spindle requires dynamic adaptation and coordination among an array of motors and crosslinkers. A new study demonstrates in vitro how the mitotic crosslinker PRC1 can tune its behavior to regulate the speed of microtubule sliding.

Automated tracking of S. pombe spindle elongation dynamics.

The mitotic spindle is a microtubule-based machine that pulls the two identical sets of chromosomes to opposite ends of the cell during cell division. The fission yeast Schizosaccharomyces pombe is an important model organism for studying mitosis due to its simple, stereotyped spindle structure and well-established genetic toolset. S.

Viscoelastic Relaxation of the Nuclear Envelope Does Not Cause the Collapse of the Spindle After Ablation in .

A large molecular machine called the mitotic spindle is responsible for accurate chromosome segregation in eukaryotic cells. The spindle consists of protein filaments known as microtubules and microtubule-associated proteins such as motors and crosslinkers, which help impart its organization. In the case of the fission yeast , these form a single bundle inside the nucleus.

Knitting Ripples.

Ripples are common in both biological systems and human clothes. Knitters have long exploited the ability of fabric to curl out of plane, by either removing or adding stitches to the material as it is created. Here, we present two knitting patterns for scarves to illustrate how ripples can arise through such additive processes.

The Spindle: Integrating Architecture and Mechanics across Scales.

The spindle segregates chromosomes at cell division, and its task is a mechanical one. While we have a nearly complete list of spindle components, how their molecular-scale mechanics give rise to cellular-scale spindle architecture, mechanics, and function is not yet clear. Recent in vitro and in vivo measurements bring new levels of molecular and physical control and shed light on this question.

Mapping Load-Bearing in the Mammalian Spindle Reveals Local Kinetochore Fiber Anchorage that Provides Mechanical Isolation and Redundancy.

Active forces generated at kinetochores move chromosomes, and the dynamic spindle must robustly anchor kinetochore fibers (k-fibers) to bear this load. The mammalian spindle bears the load of chromosome movement far from poles, but we do not know where and how-physically and molecularly-this load distributes across the spindle. In part, this is because probing spindle mechanics in live cells is difficult.

The chromokinesin Klp3a and microtubules facilitate acentric chromosome segregation.

Although poleward segregation of acentric chromosomes is well documented, the underlying mechanisms remain poorly understood. Here, we demonstrate that microtubules play a key role in poleward movement of acentric chromosome fragments generated in neuroblasts. Acentrics segregate with either telomeres leading or lagging in equal frequency and are preferentially associated with peripheral bundled microtubules.

Force on spindle microtubule minus ends moves chromosomes.

The spindle is a dynamic self-assembling machine that coordinates mitosis. The spindle's function depends on its ability to organize microtubules into poles and maintain pole structure despite mechanical challenges and component turnover. Although we know that dynein and NuMA mediate pole formation, our understanding of the forces dynamically maintaining poles is limited: we do not know where and how quickly they act or their strength and structural impact.

Single-molecule fluorescence imaging of processive myosin with enhanced background suppression using linear zero-mode waveguides (ZMWs) and convex lens induced confinement (CLIC).

Resolving single fluorescent molecules in the presence of high fluorophore concentrations remains a challenge in single-molecule biophysics that limits our understanding of weak molecular interactions. Total internal reflection fluorescence (TIRF) imaging, the workhorse of single-molecule fluorescence microscopy, enables experiments at concentrations up to about 100 nM, but many biological interactions have considerably weaker affinities, and thus require at least one species to be at micromolar or higher concentration. Current alternatives to TIRF often require three-dimensional confinement, and thus can be problematic for extended substrates, such as cytoskeletal filaments.

Future challenges in single-molecule fluorescence and laser trap approaches to studies of molecular motors.

Single-molecule analysis is a powerful modern form of biochemistry, in which individual kinetic steps of a catalytic cycle of an enzyme can be explored in exquisite detail. Both single-molecule fluorescence and single-molecule force techniques have been widely used to characterize a number of protein systems. We focus here on molecular motors as a paradigm.

Detailed tuning of structure and intramolecular communication are dispensable for processive motion of myosin VI.

Dimeric myosin VI moves processively hand-over-hand along actin filaments. We have characterized the mechanism of this processive motion by measuring the impact of structural and chemical perturbations on single-molecule processivity. Processivity is maintained despite major alterations in lever arm structure, including replacement of light chain binding regions and elimination of the medial tail.

Engineered myosin VI motors reveal minimal structural determinants of directionality and processivity.

Myosins have diverse mechanical properties reflecting a range of cellular roles. A major challenge is to understand the structural basis for generating novel functions from a common motor core. Myosin VI (M6) is specialized for processive motion toward the (-) end of actin filaments.

Rapid membrane fusion of individual virus particles with supported lipid bilayers.

Many enveloped viruses employ low-pH-triggered membrane fusion during cell penetration. Solution-based in vitro assays in which viruses fuse with liposomes have provided much of our current biochemical understanding of low-pH-triggered viral membrane fusion. Here, we extend this in vitro approach by introducing a fluorescence assay using single particle tracking to observe lipid mixing between individual virus particles (influenza or Sindbis) and supported lipid bilayers.