Synthesis of distributed systems Final report, 1 Sep. 1966 - 31 Aug. 1969

Algorithm for synthesis of distributed systems to solve circuit design problem

VAMP721 conformations unmask an extended motif for K+ channel binding and gating control

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins play a major role in membrane fusion and contribute to cell expansion, signaling, and polar growth in plants. The SNARE SYP121 of Arabidopsis thaliana that facilitates vesicle fusion at the plasma membrane also binds with, and regulates, K+ channels already present at the plasma membrane to affect K+ uptake and K+-dependent growth. Here, we report that its cognate partner VAMP721, which assembles with SYP121 to drive membrane fusion, binds to the KAT1 K+ channel via two sites on the protein, only one of which contributes to channel-gating control. Binding to the VAMP721 SNARE domain suppressed channel gating. By contrast, interaction with the amino-terminal longin domain conferred specificity on VAMP721 binding without influencing gating. Channel binding was defined by a linear motif within the longin domain. The SNARE domain is thought to wrap around this structure when not assembled with SYP121 in the SNARE complex. Fluorescence lifetime analysis showed that mutations within this motif, which suppressed channel binding and its effects on gating, also altered the conformational displacement between the VAMP721 SNARE and longin domains. The presence of these two channel-binding sites on VAMP721, one also required for SNARE complex assembly, implies a well-defined sequence of events coordinating K+ uptake and the final stages of vesicle traffic. It suggests that binding begins with VAMP721, and subsequently with SYP121, thereby coordinating K+ channel gating during SNARE assembly and vesicle fusion. Thus, our findings also are consistent with the idea that the K+ channels are nucleation points for SNARE complex assembly

Infrared emission from interstellar dust cloud with two embedded sources: IRAS 19181+1349

Mid and far infrared maps of many Galactic star forming regions show multiple peaks in close proximity, implying more than one embedded energy sources. With the aim of understanding such interstellar clouds better, the present study models the case of two embedded sources. A radiative transfer scheme has been developed to deal with an uniform density dust cloud in a cylindrical geometry, which includes isotropic scattering in addition to the emission and absorption processes. This scheme has been applied to the Galactic star forming region associated with IRAS 19181+1349, which shows observational evidence for two embedded energy sources. Two independent modelling approaches have been adopted, viz., to fit the observed spectral energy distribution (SED) best; or to fit the various radial profiles best, as a function of wavelength. Both the models imply remarkably similar physical parameters.Comment: 17 pages, 6 Figures, uses epsf.sty. To appear in Journal of Astronophysics & Astronom

Voltage-sensor transitions of the inward-rectifying K+ channel KAT1 indicate a latching mechanism biased by hydration within the voltage sensor

The Kv-like K+ channels at the plasma membrane, including the inward-rectifying KAT1 K+ channel of Arabidopsis, are important targets for manipulating K+ homeostasis in plants. Gating modification, especially, has been identified as a promising means by which to engineer plants with improved characteristics in mineral and water use. Understanding plant K+ channel gating poses several challenges, despite many similarities to that of mammalian Kv and Shaker channel models. We have used site-mutagenesis to explore residues that are thought to form two electrostatic counter-charge centers either side of a conserved Phe residue within the S2 and S3 α-helices of the voltage sensor domain (VSD) of Kv channels. Consistent with molecular dynamic simulations of KAT1, we show that the voltage dependence of the channel gate is highly sensitive to manipulations affecting these residues. Mutations of the central Phe residue favored the closed KAT1 channel, whereas mutations affecting the counter-charge centers favored the open channel. Modelling of the macroscopic current kinetics also highlighted a substantial difference between the two sets of mutations. We interpret these findings in context of the effects on hydration of amino-acid residues within the VSD and with an inherent bias of the VSD, when hydrated around a central Phe residue, to the closed state of the channel

Dual sites for SEC11 on the SNARE SYP121 implicate a binding exchange during secretory traffic

SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins facilitate vesicle traffic through their assembly in a heteromeric complex that drives membrane fusion. Much of vesicle traffic at the Arabidopsis (Arabidopsis thaliana) plasma membrane is subject to the Sec1/Munc18 protein SEC11, which, along with plasma membrane K+ channels, selectively binds with the SNARE SYP121 to regulate its assembly in complex. How SEC11 binding is coordinated with the K+ channels is poorly understood, as both SEC11 and the channels are thought to compete for the same SNARE binding site. Here, we identify a second binding motif within the N terminus of SYP121 and demonstrate that this motif affects SEC11 binding independently of the F9xRF motif that is shared with the K+ channels. This second, previously unrecognized motif is centered on residues R20R21 of SYP121 and is essential for SEC11 interaction with SYP121. Mutation of the R20R21 motif blocked vesicle traffic without uncoupling the effects of SYP121 on solute and K+ uptake associated with the F9xRF motif; the mutation also mimicked the effects on traffic block observed on coexpression of the dominant-negative SEC11Δ149 fragment. We conclude that the R20R21 motif represents a secondary site of interaction for the Sec1/Munc18 protein during the transition of SYP121 from the occluded to the open conformation that leads to SNARE complex assembly

CalibNet: Geometrically Supervised Extrinsic Calibration using 3D Spatial Transformer Networks

3D LiDARs and 2D cameras are increasingly being used alongside each other in sensor rigs for perception tasks. Before these sensors can be used to gather meaningful data, however, their extrinsics (and intrinsics) need to be accurately calibrated, as the performance of the sensor rig is extremely sensitive to these calibration parameters. A vast majority of existing calibration techniques require significant amounts of data and/or calibration targets and human effort, severely impacting their applicability in large-scale production systems. We address this gap with CalibNet: a self-supervised deep network capable of automatically estimating the 6-DoF rigid body transformation between a 3D LiDAR and a 2D camera in real-time. CalibNet alleviates the need for calibration targets, thereby resulting in significant savings in calibration efforts. During training, the network only takes as input a LiDAR point cloud, the corresponding monocular image, and the camera calibration matrix K. At train time, we do not impose direct supervision (i.e., we do not directly regress to the calibration parameters, for example). Instead, we train the network to predict calibration parameters that maximize the geometric and photometric consistency of the input images and point clouds. CalibNet learns to iteratively solve the underlying geometric problem and accurately predicts extrinsic calibration parameters for a wide range of mis-calibrations, without requiring retraining or domain adaptation. The project page is hosted at https://epiception.github.io/CalibNetComment: Appeared in the proccedings of the IEEE International Conference on Intelligent Robots and Systems (IROS) 201