SUMMARY The location, size and number of synapses critically influence the specificity and strength of neural connections. In axons, synaptic vesicle (SV) and active zone (AZ) proteins are transported by molecular motors and accumulate at discrete presynaptic loci. Little is known about the mechanisms coordinating presynaptic protein transport and deposition to achieve proper distribution of synaptic material. Here we show that SV and AZ proteins exhibit extensive co-transport and undergo frequent pauses. At the axonal and synaptic pause sites, the balance between the capture and dissociation of mobile transport packets determines the extent of presynaptic assembly. The small G protein ARL-8 inhibits assembly by promoting dissociation, while a JNK kinase pathway and AZ assembly proteins inhibit dissociation. Furthermore, ARL-8 directly binds to the UNC-104/KIF1A motor to limit the capture efficiency. Together, molecular regulation of the dichotomy between axonal trafficking and local assembly controls vital aspects of synapse formation and maintenance.

Background: Human protein-disulfide isomerase (hPDI) is a key enzyme and chaperone for protein folding. Results: Oxidation of domain aЈ releases the compact conformation of hPDI and exposes the buried substrate-binding sites facilitating its high chaperone activity. Conclusion: Oxidation of hPDI activates its chaperone activity. Significance: This study provides the first structural evidence of and mechanistic insights into the redox-regulated chaperone activity of hPDI.

SummaryPlant basic helix-loop-helix (bHLH) transcription factors play essential roles in abiotic stress tolerance. However, most bHLHs have not been functionally characterized. Here, we characterized the functional role of a bHLH transcription factor from Arabidopsis, AtbHLH112, in response to abiotic stress.AtbHLH112 is a nuclear-localized protein, and its nuclear localization is induced by salt, drought and abscisic acid (ABA). In addition, AtbHLH112 serves as a transcriptional activator, with the activation domain located at its N-terminus.In addition to binding to the E-box motifs of stress-responsive genes, AtbHLH112 binds to a novel motif with the sequence 'GG[GT]CC[GT] [GA][TA]C' (GCG-box). Gain-and loss-offunction analyses showed that the transcript level of AtbHLH112 is positively correlated with salt and drought tolerance. AtbHLH112 mediates stress tolerance by increasing the expression of P5CS genes and reducing the expression of P5CDH and ProDH genes to increase proline levels. AtbHLH112 also increases the expression of POD and SOD genes to improve reactive oxygen species (ROS) scavenging ability.We present a model suggesting that AtbHLH112 is a transcriptional activator that regulates the expression of genes via binding to their GCG-or E-boxes to mediate physiological responses, including proline biosynthesis and ROS scavenging pathways, to enhance stress tolerance.

Monitoring of bolt looseness is essential for ensuring the safety and reliability of equipment and structures with bolted connections. It is well known that tangential damping has an important influence on energy dissipation during wave propagation across the bolted joints, which require different levels of preload. In this paper, the energy dissipation generated by tangential damping of the bolted joints under different bolt preloads was modeled analytically based on fractal contact theory, which took the imperfect interface into account. A saturation value exists with the increase of the bolt preload, and the center frequency of emitted signal is demonstrated to affect the received energy significantly. Compared with previous similar studies based on experimental techniques and numerical method, the investigation presented in this paper explains the phenomenon from the inherent mechanism, and achieves the accurate quantitative monitoring of bolt looseness directly, rather than an indirect failure index. Finally, the validity of the proposed method in this paper was demonstrated with an experimental study of a bolted joint with different preload levels.

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SUN proteins are the core components of LINC complexes that span across the nuclear envelope for nuclear positioning and migration. SUN proteins contain at least one predicted coiled-coil domain preceding the SUN domain. Here, we found that the two coiled-coil domains (CC1 and CC2) of SUN2 exhibit distinct oligomeric states. CC2 is a monomer in solution. The structure of the CC2-SUN monomer revealed that CC2 unexpectedly folds as a three-helix bundle that interacts with the SUN domain and locks it in an inactive conformation. In contrast, CC1 is a trimer. The structure of the CC1 trimer demonstrated that CC1 is an imperfect coiled coil for the trimerization and activation of the SUN domain. Modulations of CC1 and CC2 dictate different oligomeric states of CC1-CC2-SUN, which is essential for LINC complex formation. Thus, the two coiled-coil domains of SUN2 act as the intrinsic dynamic regulators for controlling the SUN domain activity.

In kinesin-3, the coiled-coil 1 (CC1) can sequester the preceding neck coil (NC) for autoinhibition, but the underlying mechanism is poorly understood. Here, we determined the structures of the uninhibited motor domain (MD)-NC dimer and inhibited MD-NC-CC1 monomer of kinesin-3 KIF13B. In the MD-NC-CC1 monomer, CC1 is broken into two short helices that unexpectedly interact with both the NC and the MD. Compared with the MD-NC dimer, the CC1-mediated integration of NC and MD not only blocks the NC dimer formation, but also prevents the neck linker (NL) undocking and the ADP release from the MD. Mutations of the essential residues in the interdomain interaction interface in the MD-NC-CC1 monomer restored the MD activity. Thus, CC1 fastens the neck domain and MD and inhibits both NC and NL. This CC1-mediated lockdown of the entire neck domain may represent a paradigm for kinesin autoinhibition that could be applicable to other kinesin-3 motors.

Reinforced concrete underground pipelines are some of the most widely used types of structures in water transportation systems. Cracks and leakage are the leading causes of pipeline structural failures which directly results in economic losses and environmental hazards. In this paper, the authors propose a piezoceramic based active sensing approach to detect the cracks and the further leakage of concrete pipelines. Due to the piezoelectric properties, piezoceramic material can be utilized as both the actuator and the sensor in the active sensing approach. The piezoceramic patch, which is sandwiched between protective materials called 'smart aggregates,' can be safely embedded into concrete structures. Circumferential and axial cracks were investigated. A wavelet packet-based energy analysis was developed to distinguish the type of crack and determine the further leakage based on different stress wave energy attenuation propagated through the cracks.