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ParABS partition systems, comprising the centromere-like DNA sequence parS, the parS-binding ParB-CTPase, and the nucleoid-binding ParA-ATPase, ensure faithful segregation of bacterial chromosomes and low-copy-number plasmids. F-plasmid partition complexes containing ParBF and parSF move by generating and following a local concentration gradient of nucleoid-bound ParAF. However, the process through which ParBF activates ParAF-ATPase has not been defined. We studied CTP- and parSF-modulated ParAF–ParBF complex assembly, in which DNA-bound ParAF-ATP dimers are activated for ATP hydrolysis by interacting with two ParBF N-terminal domains. CTP or parSF enhances the ATPase rate without significantly accelerating ParAF–ParBF complex assembly. Together, parSF and CTP accelerate ParAF–ParBF assembly without further significant increase in ATPase rate. Magnetic-tweezers experiments showed that CTP promotes multiple ParBF loading onto parSF-containing DNA, generating condensed partition complex-like assemblies. We propose that ParBF in the partition complex adopts a conformation that enhances ParBF–ParBF and ParAF–ParBF interactions promoting efficient partitioning.

ParABS partition systems, comprising the centromere-like DNA sequence parS, the parS-binding ParB-CTPase and the nucleoid-binding ParA-ATPase, ensure faithful segregation of bacterial chromosomes and low-copy-number plasmids. F-plasmid partition complexes containing ParBF and parSF move by generating and following a local concentration gradient of nucleoid-bound ParAF. However, the process through which ParBF activates ParAF-ATPase has not been defined. We studied CTP- and parSF-modulated ParAF—ParBF complex assembly, in which DNA-bound ParAF-ATP dimers are activated for ATP hydrolysis by interacting with two ParBF N-terminal domains. CTP or parSF enhances the ATPase rate without significantly accelerating ParAF—ParBF complex assembly. Together, parSF and CTP accelerate ParAF—ParBF assembly without further significant increase in ATPase rate. Magnetic-tweezers experiments showed that CTP promotes multiple ParBF loading onto parSF-containing DNA, generating condensed partition complex-like assemblies. We propose that ParBF in the partition complex adopts a conformation that enhances ParBF—ParBF and ParAF—ParBF interactions promoting efficient partitioning.

Bacterial cell division requires the assembly of FtsZ protofilaments into a dynamic structure called the 'Z-ring'. The Z-ring recruits the division machinery and directs local cell wall remodeling for constriction. The organization and dynamics of protofilaments within the Z-ring coordinate local cell wall synthesis during cell constriction, but their regulation is largely unknown. The disordered C-terminal linker (CTL) region of Caulobacter crescentus FtsZ (CcFtsZ) regulates polymer structure and turnover in solution in vitro, and regulates Z-ring structure and activity of cell wall enzymes in vivo. To investigate the contributions of the CTL to the polymerization properties of FtsZ on its physiological platform, the cell membrane, we reconstituted CcFtsZ polymerization on supported lipid bilayers (SLB) and visualized polymer dynamics and structure using total internal reflection fluorescence microscopy. Unlike Escherichia coli FtsZ protofilaments that organized into large, bundled patterns, CcFtsZ protofilaments assembled into small, dynamic clusters on SLBs. Moreover, CcFtsZ lacking its CTL formed large networks of straight filament bundles that underwent slower turnover than the dynamic clusters of wildtype FtsZ. Our in vitro characterization provides novel insights into species- and CTL-dependent differences between FtsZ assembly properties that are relevant to Z-ring assembly and function on membranes in vivo.

Fabrication of high-performance thermal management materials with ultra-high thermal conductivities and low CTEs is important in electronics fields. Recently we have initiated a series of investigations to fabricate such materials and found that metal-matrix composites (MMCs) uniquely fabricated are effective in improving their thermal and strength properties. In our study, to avoid the damage of filler particle surfaces, spark plasma sintering (SPS) processing was used as a processing technique. In this review article, various kinds of filler particle dispersed MMCs are introduced and their thermal properties of composites fabricated by using various methods, such as metal infiltration and vacuum hot pressing, are shown and well documented. This review article leads to strong indications regarding fabrication techniques for making better thermal management composites. Our SPS method is compared in terms of materials qualities to those fabricated by other fabrication techniques.

• Recent investigations have converged upon a new model in which ParA-type partitions as a Brownian ratchet.

Bacterial MinD and MinE form a standing oscillatory wave which positions the cell division inhibitor MinC, that binds MinD, everywhere on the membrane except at the midpoint of the cell, ensuring midcell positioning of the cytokinetic septum. During this process MinE undergoes fold switching as it interacts with different partners. We explore the exchange dynamics between major and excited states of the MinE dimer in 3 forms using 15 N relaxation dispersion NMR: the full-length protein (6-stranded β-sheet sandwiched between 4 helices) representing the resting state; a 10-residue N-terminal deletion (Δ10) mimicking the membrane-binding competent state where the N-terminal helix is detached to interact with membrane; and N-terminal deletions of either 30 (Δ30) or 10 residues with an I24N mutation (Δ10/I24N), in which the β1-strands at the dimer interface are extruded and available to bind MinD, leaving behind a 4-stranded β-sheet. Full-length MinE samples 2 “excited” states: The first is similar to a full-length/Δ10 heterodimer; the second, also sampled by Δ10, is either similar to or well along the pathway toward the 4-stranded β-sheet form. Both Δ30 and Δ10/I24N sample 2 excited species: The first may involve destabilization of the β3- and β3′-strands at the dimer interface; changes in the second are more extensive, involving further disruption of secondary structure, possibly representing an ensemble of states on the pathway toward restoration of the resting state. The quantitative information on MinE conformational dynamics involving these excited states is crucial for understanding the oscillation pattern self-organization by MinD–MinE interaction dynamics on the membrane.

Cubic boron nitride (cBN) particle-dispersed-aluminum (Al) matrix composites were fabricated from the powder mixture composed of cBN, pure Al and Al-5mass% Si alloy in liquid and solid co-existent state by spark plasma sintering (SPS) process. Al/cBN composites were well consolidated by heating at a temperature range between 798 K and 876 K for 1.56 ks by SPS. Microstructures of the composites produced were examined by scanning electron microscopy and the reaction between the cBN particle and the Al matrix was not detected. The relative packing density of the Al/cBN composite was higher than 99 % in a volume fraction range of cBN up to 45 %. The thermal conductivity of the composite increased with increasing the cBN content in the composite in a volume fraction range of cBN between 35 and 45 vol. %. The highest thermal conductivity of 305 W/mK was obtained for Al matrix composite containing 45 vol.% cBN particles.

SummaryBacterial cell division requires the assembly of FtsZ protofilaments into a dynamic structure called the ‘Z-ring’. The Z-ring recruits the division machinery and directs local cell wall remodeling for constriction. The organization and dynamics of protofilaments within the Z-ring coordinate local cell wall synthesis during cell constriction, but their regulation is largely unknown. The disordered C-terminal linker (CTL) region ofCaulobacter crescentusFtsZ (CcFtsZ) regulates polymer structure and turnover in solutionin vitro, and regulates Z-ring structure and activity of cell wall enzymesin vivo. To investigate the contributions of the CTL to the polymerization properties of FtsZ on its physiological platform, the cell membrane, we reconstitutedCcFtsZ polymerization on supported lipid bilayers (SLB) and visualized polymer dynamics and structure using total internal reflection fluorescence microscopy. UnlikeE. coliFtsZ protofilaments that organized into large, bundled patterns,CcFtsZ protofilaments assembled into small, dynamic clusters on SLBs. Moreover,CcFtsZ lacking its CTL formed large networks of straight filament bundles that underwent slower turnover than the dynamic clusters of wildtype FtsZ. Ourin vitrocharacterization provides novel insights into species- and CTL-dependent differences between FtsZ assembly properties that are relevant to Z-ring assembly and function on membranesin vivo.

DNA segregation is a critical process for all life, and although there is a relatively good understanding of eukaryotic mitosis, the mechanism in bacteria remains unclear. The small size of a bacterial cell and the number of factors involved in its subcellular organization make it difficult to study individual systems under controlled conditions in vivo. We developed a cell-free technique to reconstitute and visualize bacterial ParA-mediated segregation systems. Our studies provide direct evidence for a mode of transport that does not use a classical cytoskeletal filament or motor protein. Instead, we demonstrate that ParA-type DNA segregation systems can establish a propagating ParA ATPase gradient on the nucleoid surface, which generates the force required for the directed movement of spatially confined cargoes, such as plasmids or large organelles, and distributes multiple cargos equidistant to each other inside cells. Here we present the critical principles of our diffusion-ratchet model of ParA-mediated transport and expand on the mathematically derived chemophoresis force using experimentally-determined biochemical and cellular parameters.

This paper describes the development of a metal-core piezoelectric ceramics fiber/aluminum composite using a low cost hollow fiber. To develop this composite, this study applied a new fabrication process to fill the Al-Cu liquid phase alloy produced during the interphase forming/bonding (IF/B) method in the hollows. The IF/B method is a sophisticated transient liquid phase (TLP) bonding method developed to enable embedding fragile functional fibers at low temperature and low pressure in a short time period by using the eutectic reaction of inserting material and matrix. To increase the filling rate as well as removing the voids and the excessive Al-Cu alloy that remained in the matrix, the effects of the hot pressing temperature, the pressure, and the period were optimized in experiments. Under a hot pressing temperature of 873 K, a pressure of 2.2 MPa, and a period for 3.0 ks, the filling rate attained 99% without harmful chemical reactions between the fiber and the matrix, and the voids were successfully removed. In addition, it was clarified that the Al-Cu alloy core and the aluminum matrix worked as electrodes and that the embedded fiber could generate a relatively high output voltage, as well as the conventional metal-core piezoelectric ceramics fiber.

Bacterial plasmids are extrachromosomal DNA that provides selective advantages for bacterial survival. Plasmid partitioning can be remarkably robust. For high-copy-number plasmids, diffusion ensures that both daughter cells inherit plasmids after cell division. In contrast, most low-copy-number plasmids need to be actively partitioned by a conserved tripartite ParA-type system. ParA is an ATPase that binds to chromosomal DNA; ParB is the stimulator of the ParA ATPase and specifically binds to the plasmid at a centromere-like site, parS. ParB stimulation of the ParA ATPase releases ParA from the bacterial chromosome, after which it takes a long time to reset its DNA-binding affinity. We previously demonstrated in vitro that the ParA system can exploit this biochemical asymmetry for directed cargo transport. Multiple ParA-ParB bonds can bridge a parS-coated cargo to a DNA carpet, and they can work collectively as a Brownian ratchet that directs persistent cargo movement with a ParA-depletion zone trailing behind. By extending this model, we suggest that a similar Brownian ratchet mechanism recapitulates the full range of actively segregated plasmid motilities observed in vivo. We demonstrate that plasmid motility is tuned as the replenishment rate of the ParA-depletion zone progressively increases relative to the cargo speed, evolving from diffusion to pole-to-pole oscillation, local excursions, and, finally, immobility. When the plasmid replicates, the daughters largely display motilities similar to that of their mother, except that when the single-focus progenitor is locally excursive, the daughter foci undergo directed segregation. We show that directed segregation maximizes the fidelity of plasmid partition. Given that local excursion and directed segregation are the most commonly observed modes of plasmid motility in vivo, we suggest that the operation of the ParA-type partition system has been shaped by evolution for high fidelity of plasmid segregation.

SUMMARYThe MinD and MinE proteins ofEscherichia coliself-organize into a standing-wave oscillator on the membrane to help align division at mid-cell. When unleashed from cellular confines, we find that MinD and MinE form a wide spectrum of patterns on artificial bilayers - static amoebas, traveling waves, traveling mushrooms, and bursts with standing-wave dynamics. We recently focused our cell-free studies on bursts because their dynamics closely resemble those foundin vivo. The data unveiled a patterning mechanism largely governed by MinE regulation of MinD interaction with membrane. We proposed that the MinD to MinE ratio on the membrane acts as a toggle switch between MinE-stimulated recruitment or release of MinD from the membrane. Here we provide data that further refines and extends our model that explains the remarkable spectrum of patterns supported by these two ‘simple’ proteins.

Diamond-particle-dispersed aluminum (Al) matrix composites consisting of monomodal and bimodal diamond particles were fabricated in spark plasma sintering process, where the mixture of diamond, pure Al and Al–5 mass% Si alloy powders were consolidated in liquid and solid co-existent state. Microstructures and thermal properties of the composites fabricated in such a way were investigated and the monomodal and bimodal diamond particle effect was evaluated on the thermal properties of the composites. The composites can be well consolidated in a temperature range between 773 K and 878 K and scanning electron microscopy detects no reaction product at the interface between the diamond particle and the Al matrix. Relative packing density of the composite containing monomodal diamond particles decreased from 99.1% to 87.4% with increasing volume fraction of diamond between 50% and 60%, whereas that of the composite containing bimodal diamond particles was higher than 99% in a volume fraction of diamond up to 65%. The thermal conductivity of the composite containing bimodal diamond particles was higher than that of the composite containing monomodal diamond particles in a volume fraction of diamond higher than 60%. The coefficients of thermal expansion (CTEs) of the diamond-particle-dispersed Al–matrix composites fall in the upper line of Kerner model, indicating good bonding between the diamond particle and the Al matrix in the composite. The thermal conductivity of the composite containing 70 vol.% bimodal diamond particles was 578 W/m K and its CTE was 6.72 × 10 −6 at R.T.

Summary The E. coli Min system forms a cell-pole-to-cell-pole oscillator that positions the divisome at mid-cell. The MinD ATPase binds the membrane and recruits the cell division inhibitor MinC. MinE interacts with and releases MinD (and MinC) from the membrane. The chase of MinD by MinE creates the in vivo oscillator that maintains a low level of the division inhibitor at mid-cell. In vitro reconstitution and visualization of Min proteins on a supported lipid bilayer has provided significant advances in understanding Min patterns in vivo. Here we studied the effects of flow, lipid composition, and salt concentration on Min patterning. Flow and no-flow conditions both supported Min protein patterns with somewhat different characteristics. Without flow, MinD and MinE formed spiraling waves. MinD and, to a greater extent MinE, have stronger affinities for anionic phospholipid. MinD-independent binding of MinE to anionic lipid resulted in slower and narrower waves. MinE binding to the bilayer was also more susceptible to changes in ionic strength than MinD. We find that modulating protein diffusion with flow, or membrane binding affinities with changes in lipid composition or salt concentration, can differentially affect the retention time of MinD and MinE, leading to spatiotemporal changes in Min patterning.

Diamond-particle-dispersed aluminum (Al) matrix composites consisting of monomodal and bimodal diamond particles were fabricated in spark plasma sintering process, where the mixture of diamond, pure Al and Al-5mass% Si alloy powders were consolidated in liquid and solid co-existent state. Microstructures and thermal properties of the composites fabricated in such a unique way were investigated and the bimodal and monomodal diamond particle effect was evaluated on the thermal properties of the composites. The composites can be well consolidated in a temperature range between 773 K and 878 K and scanning electron microscopy detects no reaction product at the interface between the diamond particle and the Al matrix. Relative packing density of the composite containing monomodal diamond particles decreased from 99.1% to 87.4% with increasing volume fraction of diamond between 50% and 60%, whereas that of the composite containing bimodal diamond particles was higher than 99% in a volume fraction of diamond up to 65%. The thermal conductivity of the composite containing bimodal diamond particles was higher than that of the composite containing monomodal diamond particles in a volume fraction of diamond higher than 60% and the thermal conductivity of the composite containing 70 vol.% bimodal diamond particles was 578 W/mK at R.T..

The faithful segregation of duplicated genetic material into daughter cells is critical to all organisms. In many bacteria, the segregation of chromosomes involves transport of “centromere-like” loci over the main body of the chromosome, the nucleoid, mediated by a two-protein partition system: a nonspecific DNA-binding ATPase, ParA, and an ATPase stimulator, ParB, which binds to the centromere-like loci. These systems have previously been proposed to function through a filament-based mechanism, analogous to actin- or microtubule-based movement. Here, we reconstituted the F-plasmid partition system using a DNA-carpeted flow cell as an artificial nucleoid surface and magnetic beads coated with plasmid partition complexes as surface-confined cargo. This minimal system recapitulated directed cargo motion driven by a surface ATPase gradient that propagated with the cargo. The dynamics are consistent with a diffusion-ratchet model, whereby the cargo dynamically establishes, and interacts with, a concentration gradient of the ATPase. A chemophoresis force ensues as the cargo perpetually chases the ATPase gradient, allowing the cargo to essentially “surf” the nucleoid on a continuously traveling wave of the ATPase. Demonstration of this non–filament-based motility mechanism in a biological context establishes a distinct class of motor system used for the transport and positioning of large cellular cargo.

The Escherichia coli Min system self-organizes into a cell-pole to cell-pole oscillator on the membrane to prevent divisions at the cell poles. Reconstituting the Min system on a lipid bilayer has contributed to elucidating the oscillatory mechanism. However, previous in vitro patterns were attained with protein densities on the bilayer far in excess of those in vivo and failed to recapitulate the standing wave oscillations observed in vivo. Here we studied Min protein patterning at limiting MinD concentrations reflecting the in vivo conditions. We identified “burst” patterns—radially expanding and imploding binding zones of MinD, accompanied by a peripheral ring of MinE. Bursts share several features with the in vivo dynamics of the Min system including standing wave oscillations. Our data support a patterning mechanism whereby the MinD-to-MinE ratio on the membrane acts as a toggle switch: recruiting and stabilizing MinD on the membrane when the ratio is high and releasing MinD from the membrane when the ratio is low. Coupling this toggle switch behavior with MinD depletion from the cytoplasm drives a self-organized standing wave oscillator.

Spatial organization and active transportation of organelles are essential processes for cellular division. This is true even in bacteria, which were once thought too simple to require much internal organization. The ubiquitous deviant Walker ATPase family of proteins has been shown to be involved in systems important for spatial coordination in bacteria, the two most highly studied examples of which are the DNA partitioning Par system and the divisome positioning Min system. Both these systems contain the eponymous deviant Walker A ATPase (ParA or MinD), a protein which stimulates the ATPase's hydrolysis activity (ParB or MinE) and a cargo to be transported or localized (a “centromere” DNA site bound by ParB or the divisome inhibitor MinC bound to MinD). Despite the similarities of their components these systems appear to behave quite differently in vivo: the Par system segregates large clusters of ParB bound DNA cargo into sister cells during division by displacing ParA non-specifically bound to the nucleoid whereas the Min proteins bind the inner membrane and are seen to oscillate from one cell pole to the other. These differences may arise due to the nature of the binding surface (DNA vs lipid) or the nature of the cargo (a large cargo bound a large number of stimulator protein molecules in Par vs small protein molecules individually bound to membrane-bound MinD dimers). Here we use TIRF microscopy to demonstrate that the Min system is capable of transporting a magnetic bead (a large cargo) along a supported lipid bilayer in a manner strikingly similar to our previous reconstitution of the Par system. This demonstrates that despite their clear differences in their system dynamics under conditions for their natural functions, Par and Min systems operate based on common underlying mechanistic principles.

Summary The ParA family of ATPases is responsible for transporting bacterial chromosomes, plasmids and large protein machineries. ParAs pattern the nucleoid in vivo, but how patterning functions or is exploited in transport is of considerable debate. Here we discuss the process of self-organization into patterns on the bacterial nucleoid and explore how it relates to the molecular mechanism of ParA action. We review ParA-mediated DNA partition as a general mechanism of how ATP-driven protein gradients on biological surfaces can result in spatial organization on a mesoscale. We also discuss how the nucleoid acts as a formidable diffusion barrier for large bodies in the cell, and make the case that the ParA family evolved to overcome the barrier by exploiting the nucleoid as a matrix for movement.

Silicon carbide (SiC)-particle-dispersed-aluminum (Al) matrix composites were fabricated in a unique fabrication method, where the powder mixture of SiC, pure Al and Al–5mass% Si alloy was uniquely designed to form continuous solid–liquid co-existent state during spark plasma sintering (SPS) process. Composites fabricated in such a way can be well consolidated by heating during SPS processing in a temperature range between 798 K and 876 K for a heating duration of 1.56 ks. Microstructures of the composites thus fabricated were examined by scanning electron microscopy and no reaction was detected at the interface between the SiC particle and the Al matrix. The relative packing density of the Al–matrix composite containing SiC was higher than 99% in a volume fraction range of SiC between 40% and 55%. Thermal conductivity of the composite increased with increasing the SiC content in the composite at a SiC fraction range between 40 vol.% and 50 vol.%. The highest thermal conductivity was obtained for Al–50 vol.% SiC composite and reached 252 W/mK. The coefficient of thermal expansion of the composites falls in the upper line of Kerner’s model, indicating strong bonding between the SiC particle and the Al matrix in the composite.

Aluminum nitride-particle-dispersed aluminum–matrix composites were fabricated in a unique fabrication method, where the powder mixture of AlN, pure Al and Al–5 mass%Si alloy was uniquely designed to form continuous solid–liquid co-existent state during spark plasma sintering (SPS) process. Composites fabricated in such a way can be well consolidated by heating during SPS processing in a temperature range between 798 K and 876 K for a heating duration of 1.56 ks. Microstructures of the composites thus fabricated were examined by scanning electron microscopy and no reaction product was detected at the interface between the AlN particle and the Al matrix. The relative packing density of the Al/AlN composite was almost 100% when volume fraction of AlN is between 40% and 60%. Thermal conductivity of the composite was higher than 180 W/mK at an AlN fraction range between 40 and 65 vol.%, approximately 90% of the theoretical thermal conductivity estimated by Maxwell–Eucken’s model. The coefficient of thermal expansion of the composite falls in the upper line of Kerner’s model, indicating strong bonding between the AlN particle and the Al matrix in the composite.

Diamond-particle-dispersed silver (Ag) matrix composite was fabricated in a unique fabrication method, where solid–liquid coexistent state of the powder mixture of diamond, pure Ag and pure Si was designed to create during spark plasma sintering (SPS) process. The composite is well consolidated in a temperature range between 1113 K and 1188 K and no reaction was detected by scanning electron microscopy at the interface between diamond particles and the Ag matrix. The relative packing density of the diamond–Ag composite fabricated was 95–97% in a volume fraction range of diamond between 40% and 50%. The thermal conductivity of the diamond–Ag composite containing 50 vol.% (v/o) diamond reached 717 W/mK, approximately 80% the theoretical thermal conductivity calculated by Maxwell–Eucken’s equation. This result suggests that the solid–liquid co-existent state during SPS consolidation is very effective not only for rapid densification of the composite but also for producing strong bonding between the diamond particles and the Ag matrix.

Integration of viral DNA into the host genome is an essential step in retroviral replication that is mediated by a stable nucleoprotein complex comprising a tetramer of integrase bridging the two ends of the viral DNA in a stable synaptic complex (SSC) or intasome. Assembly of HIV-1 intasomes requires several hundred base pairs of nonspecific internal DNA in addition to the terminal viral DNA sequence that is protected in footprinting experiments. We find that only one of the viral DNA ends in the intasome requires long-nonspecific internal DNA for intasome assembly. Although intasomes are unstable in solution when the nonspecific internal DNA is cut off after assembly, they are stable in agarose gels. These complexes are indistinguishable from SSCs with nonspecific internal DNA in Forster resonance energy transfer (FRET) experiments suggesting the interactions with the viral DNA and integrase tetramer are the same regardless of the presence of nonspecific internal DNA. We discuss models of how the internal DNA contributes to intasome assembly and stability. FRET is exquisitely sensitive to the distance between the fluorophores and given certain assumptions can be translated to distance measurements. We anticipated that a set of such distance constraints would provide a map of the DNA path within the intasome. In reality, the constraints we could impose from the FRET data were quite weak allowing a wide envelope for the possible path. We discuss the difficulties of converting the FRET signal to absolute distance within nucleoprotein complexes.

Diamond-particle-dispersed-aluminum (Al) matrix composites were fabricated in continuous solid-liquid co-existent state by spark plasma sintering (SPS) process from the mixture of diamond powders, pure Al powders and Al-5mass%Si alloy powders. The microstructures and thermal conductivities of the composites fabricated were examined. These composites were well consolidated by heating at a temperature range between 798K and 876K for 1.56ks during SPS process. No reaction at the interface between the diamond particle and the Al matrix was observed by scanning electron microscopy for the composites fabricated under the sintering conditions employed in the present study. The relative packing density of the diamond-Al composite fabricated was 99% or higher in a volume fraction range of diamond between 45% and 50%. Thermal conductivity of the diamond-Al composite containing 50 vol.% diamond reached 552W/mK, approximately 95% the theoretical thermal conductivity estimated using Maxwell-Eucken’s equation.

The segregation of DNA before cell division is essential for faithful genetic inheritance. In many bacteria, segregation of low-copy number plasmids involves an active partition system composed of a nonspecific DNA-binding ATPase, ParA, and its stimulator protein ParB. The ParA/ParB system drives directed and persistent movement of DNA cargo both in vivo and in vitro. Filament-based models akin to actin/microtubule-driven motility were proposed for plasmid segregation mediated by ParA. Recent experiments challenge this view and suggest that ParA/ParB system motility is driven by a diffusion ratchet mechanism in which ParB-coated plasmid both creates and follows a ParA gradient on the nucleoid surface. However, the detailed mechanism of ParA/ParB-mediated directed and persistent movement remains unknown. Here, we develop a theoretical model describing ParA/ParB-mediated motility. We show that the ParA/ParB system can work as a Brownian ratchet, which effectively couples the ATPase-dependent cycling of ParA-nucleoid affinity to the motion of the ParB-bound cargo. Paradoxically, this resulting processive motion relies on quenching diffusive plasmid motion through a large number of transient ParA/ParB-mediated tethers to the nucleoid surface. Our work thus sheds light on an emergent phenomenon in which nonmotor proteins work collectively via mechanochemical coupling to propel cargos-an ingenious solution shaped by evolution to cope with the lack of processive motor proteins in bacteria.

Diamond particle dispersed aluminum (Al) matrix composites were fabricated in solid–liquid coexistent state from the powder mixture of diamond, pure Al and 5056-type Al–Mg alloy by spark plasma sintering (SPS) process. Microstructures and thermal conductivities of the composites fabricated were examined. These composites can be well consolidated at 813 K for 0.6 k s and scanning electron microscopy detects no reaction at the interface between the diamond particle and the Al matrix. The relative packing density of the diamond-Al composite fabricated was 97% or higher in a volume fraction range of diamond between 25.5 and 45.5%. The thermal conductivity of the Al matrix composite containing 45.5 vol.% diamond reached 403 W/mK, approximately 76% the theoretical thermal conductivity estimated by Maxwell–Eucken’s equation.

Diamond-particle-dispersed-aluminum (Al) matrix composites were fabricated in a unique fabrication method where continuous solid-liquid coexistent state of the powder mixture of diamond, pure Al and Al-5mass%Si alloy was designed during spark plasma sintering (SPS) process. Microstructures and thermal properties of the composites fabricated in such a way were investigated. The composites can be well consolidated in the temperature range between 798 K and 876 K and scanning electron microscopy detects no reaction at the interface between the diamond particle and the Al matrix. The relative packing density of the diamond-Al composite fabricated was 99 % or higher in a volume fraction range of diamond between 35 and 50 %. The thermal conductivity of the diamond-Al composite containing 50 vol.% diamond reached 552 W/mK, higher than 95 % the theoretical thermal conductivity calculated by Maxwell-Eucken's equation. The coefficient of thermal expansion of the composites falls in the upper line of Kerner's model, indicating strong bonding between the diamond particle and the Al matrix in the composite. Based on the experimental results obtained in the present study, densification process in continuous solid-liquid co-existent state is considered to be a quite effective methodology for enhancing performance of heat dissipative metal matrix composites.

SiC-particle-dispersed-aluminum (Al) matrix composites were fabricated in solid-liquid co-existent state by Spark Plasma Sintering (SPS) process from the mixture of SiC powders, Al powders and Al-5mass%Si powders. The microstructures and thermal conductivities of the composites fabricated were examined. These composites were all well consolidated by heating at a temperature range between 798 K and 876 K for 1.56 ks during SPS process. No reaction at the interface between the SiC particle and the Al matrix was observed by scanning electron microscopy for the composites fabricated under the sintering conditions employed in the present study. The relative packing density of the Al/SiC composite fabricated was higher than 99 % in a volume fraction range of SiC between 40 % and 55 %. Thermal conductivity of the Al/SiC composite increased with increasing the SiC content in the composite in a volume fraction range between 40 and 50 vol.%. The highest thermal conductivity was obtained for Al-50vol.%SiC composite and reached 252 W/mK. The coefficient of thermal expansion of the composites falls in the upper line of Kerner's model, indicating strong bonding between the SiC particle and the Al matrix in the composite.

DNA transposons integrate into host chromosomes with limited target sequence specificity. Without mechanisms to avoid insertion into themselves, transposons risk self-destruction. Phage Mu avoids this problem by transposition immunity, involving MuA-transposase and MuB ATP-dependent DNA-binding protein. MuB-bound DNA acts as an efficient transposition target, but MuA clusters bound to Mu DNA ends activate the MuB-ATPase and dissociate MuB from their neighborhood before target site commitment, making the regions near Mu ends a poor target. This MuA-cluster-MuB interaction requires formation of DNA loops between the MuA- and the MuB-bound DNA sites. At early times, MuB clusters are disassembled via loops with smaller average size, and at later times, MuA clusters find distantly located MuB clusters by forming loops with larger average sizes. We demonstrate that iterative loop formation/disruption cycles with intervening diffusional steps result in larger DNA loops, leading to preferential insertion of the transposon at sites distant from the transposon ends.

The MinD and MinE proteins of Escherichia coli self-organize into a standing-wave oscillator on the membrane to help align division at mid-cell. When unleashed from cellular confines, MinD and MinE form a spectrum of patterns on artificial bilayers - static amoebas, traveling waves, traveling mushrooms, and bursts with standing-wave dynamics. We recently focused our cell-free studies on bursts because their dynamics recapitulate many features of Min oscillation observed in vivo. The data unveiled a patterning mechanism largely governed by MinE regulation of MinD interaction with membrane. We proposed that the MinD to MinE ratio on the membrane acts as a toggle switch between MinE-stimulated recruitment and release of MinD from the membrane. In this review, we summarize cell-free data on the Min system and expand upon a molecular mechanism that provides a biochemical explanation as to how these two ‘simple’ proteins can form the remarkable spectrum of patterns.

Min proteins of the Escherichia coli cell division system oscillate between the cell poles in vivo. In vitro on a solid-surface supported lipid bilayer, these proteins exhibit a number of interconverting modes of collective ATP-driven dynamic pattern formation including not only the previously described propagating waves, but also near uniformity in space surface concentration oscillation, propagating filament like structures with a leading head and decaying tail and moving and dividing amoeba-like structures with sharp edges. We demonstrate that the last behavior most closely resembles in vivo system behavior. The simple reaction-diffusion models previously proposed for the Min system fail to explain the results of the in vitro self-organization experiments. We propose the hypotheses that initiation of MinD binding to the surface is controlled by counteraction of initiation and dissociation complexes; the binding of MinD/E is stimulated by MinE and involves polymerization-depolymerization dynamics; polymerization of MinE over MinD oligomers triggers dynamic instability leading to detachment from the membrane. The physical properties of the lipid bilayer are likely to be one of the critical determinants of certain aspects of the dynamic patterns observed.

Diamond-particle-dispersed copper (Cu) matrix composites were fabricated from Cu-coated diamond particles by spark plasma sintering (SPS) process, and the microstructure and thermal properties of the composites fabricated were examined. These composites can well be consolidated in a temperature range between 973K and 1173K and scanning electron microscopy detects no reaction at the interface between the diamond particle and the Cu matrix. The relative packing density of the diamond-Cu composite increases with increasing sintering temperature and holding time, reaching 99.2% when sintered at a temperature of 1173K for a holding time of 2.1ks. Thermal conductivity of the diamond-Cu composite containing 43.2 vol. % diamond increases with increasing relative packing density, reaching a maximum (654W/mK) at a relative packing density of 99.2%. This thermal conductivity is 83% the theoretical value estimated by Maxwell-Eucken equation. The coefficient of thermal expansion of the composites falls in the upper line of Kerner’s model, indicating strong bonding between the diamond particle and the Cu matrix in the composite.

Capabilities of synthesizing new structural and functional materials by SPS processing were indicated by exemplifying the synthesis of nano-structured alumina with high bending strength or high transparency, Al/diamond composites with high thermal conductivity and zirconia(3Y)/ SUS410L FGM. In the synthesis of alumina, the bending strength of more than 720MPa was attained by choosing suitable SPS conditions. It was also indicated that SPS processing could easily synthesize Al/diamond composites with high thermal conductivity of more than 400W/(m･K), suggesting elaborate control of interface between Al and diamond in SPS consolidation. Further, zirconia(3Y)/SUS410L FGM could easily be fabricated by SPS. Mechanical weakness in the zirconia(3Y)-rich layers of the FGM was shown from the analysis of stress state based on Raman scattering method. It is suggested that the designing of the layer staking in FGM based on the Raman scattering analysis is effective for the improvement of the weakness in the FGM.