Structural and functional analysis of two universal stress proteins YdaA and YnaF from Salmonella typhimurium: possible roles in microbial stress tolerance

In many organisms ``Universal Stress Proteins'' CUSPS) are induced in response to a variety of environmental stresses. Here we report the structures of two USPs, YnaF and YdaA from Salmonella typhimurium determined at 1.8 angstrom and 2.4 angstrom resolutions, respectively. YnaF consists of a single USP domain and forms a tetrameric organization stabilized by interactions mediated through chloride ions. YdaA is a larger protein consisting of two tandem USP domains. Two protomers of YdaA associate to form a structure similar to the YnaF tetramer. YdaA showed ATPase activity and an ATP binding motif G-2X-G-9X-G(S/T/N) was found in its C-terminal domain. The residues corresponding to this motif were not conserved in YnaF although YnaF could bind ATP. However, unlike YdaA, YnaF did not hydrolyse ATP in vitro. Disruption of interactions mediated through chloride ions by selected mutations converted YnaF into an ATPase. Residues that might be important for ATP hydrolysis could be identified by comparing the active sites of native and mutant structures. Only the C-terminal domain of YdaA appears to be involved in ATP hydrolysis. The structurally similar N-terminal domain was found to bind a zinc ion near the segment equivalent to the phosphate binding loop of the C-terminal domain. Mass spectrometric analysis showed that YdaA might bind a ligand of approximate molecular weight 800 daltons. Structural comparisons suggest that the ligand, probably related to an intermediate in lipid A biosynthesis, might bind at a site close to the zinc ion. Therefore, the N-terminal domain of YdaA binds zinc and might play a role in lipid metabolism. Thus, USPs appear to perform several distinct functions such as ATP hydrolysis, altering membrane properties and chloride sensing. (C) 2015 Elsevier Inc. All rights reserved.

Timer-Based Distributed Node Selection Scheme Exploiting Power Control and Capture

Opportunistic selection in multi-node wireless systems improves system performance by selecting the ``best'' node and by using it for data transmission. In these systems, each node has a real-valued local metric, which is a measure of its ability to improve system performance. Our goal is to identify the best node, which has the largest metric. We propose, analyze, and optimize a new distributed, yet simple, node selection scheme that combines the timer scheme with power control. In it, each node sets a timer and transmit power level as a function of its metric. The power control is designed such that the best node is captured even if. other nodes simultaneously transmit with it. We develop several structural properties about the optimal metric-to-timer-and-power mapping, which maximizes the probability of selecting the best node. These significantly reduce the computational complexity of finding the optimal mapping and yield valuable insights about it. We show that the proposed scheme is scalable and significantly outperforms the conventional timer scheme. We investigate the effect of. and the number of receive power levels. Furthermore, we find that the practical peak power constraint has a negligible impact on the performance of the scheme.

Metallophosphoesterases: structural fidelity with functional promiscuity

Calcineurin-like metallophosphoesterases (MPEs) form a large superfamily of binuclear metal-ion-centre-containing enzymes that hydrolyse phosphomono-, phosphodi-or phosphotri-esters in a metal-dependent manner. The MPE domain is found in Mre11/SbcD DNA-repair enzymes, mammalian phosphoprotein phosphatases, acid sphingomyelinases, purple acid phosphatases, nucleotidases and bacterial cyclic nucleotide phosphodiesterases. Despite this functional diversity, MPEs show a remarkably similar structural fold and active-site architecture. In the present review, we summarize the available structural, biochemical and functional information on these proteins. We also describe how diversification and specialization of the core MPE fold in various MPEs is achieved by amino acid substitution in their active sites, metal ions and regulatory effects of accessory domains. Finally, we discuss emerging roles of these proteins as non-catalytic protein-interaction scaffolds. Thus we view the MPE superfamily as a set of proteins with a highly conserved structural core that allows embellishment to result in dramatic and niche-specific diversification of function.

Iron(III) Complexes of a Pyridoxal Schiff Base for Enhanced Cellular Uptake with Selectivity and Remarkable Photocytotoxicity

Iron(III) complexes of pyridoxal (vitamin B6, VB6) or salicylaldehyde Schiff bases and modified dipicolylamines, namely, Fe(B)(L)](NO3) (15), where B is phenyl-N,N-bis((pyridin-2-yl)methyl)methanamine (phbpa in 1), (anthracen-9-yl)-N,N-bis((pyridin-2-yl)methyl)methanamine (anbpa in 2, 4) and (pyren-1-yl)-N,N-bis((pyridin-2-yl)methyl)methanamine (pybpa in 3, 5) (H2L1 is 3-hydroxy-5-(hydroxymethyl)-4-(((2-hydroxyphenyl)imino)methyl)-2-methylp yridine (13) and H2L2 is 2-(2-hydroxyphenyl-imino)methyl]phenol), were prepared and their uptake in cancer cells and photocytotoxicity were studied. Complexes 4 and 5, having a non-pyridoxal Schiff base, were prepared to probe the role of the pyridoxal group in tumor targeting and cellular uptake. The PF6 salt (1a) of complex 1 is structurally characterized. The complexes have a distorted six-coordinate FeN4O2 core where the metal is in the +3 oxidation state with five unpaired electrons. The complexes display a ligand to metal charge transfer band near 520 and 420 nm from phenolate to the iron(III) center. The photophysical properties of the complexes are explained from the time dependent density functional theory calculations. The redox active complexes show a quasi-reversible Fe(III)/Fe(II) response near -0.3 V vs saturated calomel electrode. Complexes 2 and 3 exhibit remarkable photocytotoxicity in various cancer cells with IC50 values ranging from 0.4 to 5 mu M with 10-fold lower dark toxicity. The cell death proceeded by the apoptotic pathway due to generation of reactive oxygen species upon light exposure. The nonvitamin complexes 4 and 5 display 3-fold lower photocytotoxicity compared to their VB6 analogues, possibly due to preferential and faster uptake of the vitamin complexes in the cancer cells. Complexes 2 and 3 show significant uptake in the endoplasmic reticulum, while complexes 4 and 5 are distributed throughout the cells without any specific localization pattern.

Molecular and Functional Characterization of RecD, a Novel Member of the SF1 Family of Helicases, from Mycobacterium tuberculosis

Background: The heterotrimeric M. tuberculosis RecBCD complex, or each of its individual subunits, remains uncharacterized. Results: MtRecD exists as a homodimer in solution, catalyzes ssDNA-dependent ATP hydrolysis, unwinding of DNA replication/recombination intermediates, and interacts with RecA. Conclusion: MtRecD possesses strong 5 3- and weak 3 5-helicase activities. Significance: These findings provide insights into the mechanism underlying DSB repair and homologous recombination in mycobacteria. The annotated whole-genome sequence of Mycobacterium tuberculosis revealed the presence of a putative recD gene; however, the biochemical characteristics of its encoded protein product (MtRecD) remain largely unknown. Here, we show that MtRecD exists in solution as a stable homodimer. Protein-DNA binding assays revealed that MtRecD binds efficiently to single-stranded DNA and linear duplexes containing 5 overhangs relative to the 3 overhangs but not to blunt-ended duplex. Furthermore, MtRecD bound more robustly to a variety of Y-shaped DNA structures having 18-nucleotide overhangs but not to a similar substrate containing 5-nucleotide overhangs. MtRecD formed more salt-tolerant complexes with Y-shaped structures compared with linear duplex having 3 overhangs. The intrinsic ATPase activity of MtRecD was stimulated by single-stranded DNA. Site-specific mutagenesis of Lys-179 in motif I abolished the ATPase activity of MtRecD. Interestingly, although MtRecD-catalyzed unwinding showed a markedly higher preference for duplex substrates with 5 overhangs, it could also catalyze significant unwinding of substrates containing 3 overhangs. These results support the notion that MtRecD is a bipolar helicase with strong 5 3 and weak 3 5 unwinding activities. The extent of unwinding of Y-shaped DNA structures was approximate to 3-fold lower compared with duplexes with 5 overhangs. Notably, direct interaction between MtRecD and its cognate RecA led to inhibition of DNA strand exchange promoted by RecA. Altogether, these studies provide the first detailed characterization of MtRecD and present important insights into the type of DNA structure the enzyme is likely to act upon during the processes of DNA repair or homologous recombination.

Temperature compensation in CNT-composite distributed strain sensors

A temperature compensation method is proposed for CNT-composite strain sensors. CNT-composite sensors are fabricated on an elastic polymer substrate having known thermo-mechanical properties to introduce thermo-mechanical strain and further calibration of the sensor. Strain is induced on the sensor by bending the substrate as a cantilever configuration. Response of the sensor is measured using a bridge circuit method. Induced strain in the beam is determined using beam theory. The sensors are characterized for different CNT concentrations and at various temperatures. A model based temperature compensation scheme is proposed and verified experimentally. The result proves the ability of CNT-nanocomposite strain sensors to be used under varying temperature applications. A method is proposed to determine the strain and temperature simultaneously. The CNT sensors are simple to fabricate in complex patterns with excellent repeatability and do not require bonding layer.

Functional characterization of heat-shock protein 90 from Oryza sativa and crystal structure of its N-terminal domain

Heat-shock protein 90 (Hsp90) is an ATP-dependent molecular chaperone that is essential for the normal functioning of eukaryotic cells. It plays crucial roles in cell signalling, cell-cycle control and in maintaining proteome integrity and protein homeostasis. In plants, Hsp90s are required for normal plant growth and development. Hsp90s are observed to be upregulated in response to various abiotic and biotic stresses and are also involved in immune responses in plants. Although there are several studies elucidating the physiological role of Hsp90s in plants, their molecular mechanism of action is still unclear. In this study, biochemical characterization of an Hsp90 protein from rice (Oryza sativa; OsHsp90) has been performed and the crystal structure of its N-terminal domain (OsHsp90-NTD) was determined. The binding of OsHsp90 to its substrate ATP and the inhibitor 17-AAG was studied by fluorescence spectroscopy. The protein also exhibited a weak ATPase activity. The crystal structure of OsHsp90-NTD was solved in complex with the nonhydrolyzable ATP analogue AMPPCP at 3.1 angstrom resolution. The domain was crystallized by cross-seeding with crystals of the N-terminal domain of Hsp90 from Dictyostelium discoideum, which shares 70% sequence identity with OsHsp90-NTD. This is the second reported structure of a domain of Hsp90 from a plant source.

Insights into the Functional Roles of N-Terminal and C-Terminal Domains of Helicobacter pylori DprA

DNA processing protein A (DprA) plays a crucial role in the process of natural transformation. This is accomplished through binding and subsequent protection of incoming foreign DNA during the process of internalization. DprA along with Single stranded DNA binding protein A (SsbA) acts as an accessory factor for RecA mediated DNA strand exchange. H. pylori DprA (HpDprA) is divided into an N-terminal domain and a C-terminal domain. In the present study, individual domains of HpDprA have been characterized for their ability to bind single stranded (ssDNA) and double stranded DNA (dsDNA). Oligomeric studies revealed that HpDprA possesses two sites for dimerization which enables HpDprA to form large and tightly packed complexes with ss and dsDNA. While the N-terminal domain was found to be sufficient for binding with ss or ds DNA, C-terminal domain has an important role in the assembly of poly-nucleoprotein complex. Using site directed mutagenesis approach, we show that a pocket comprising positively charged amino acids in the N-terminal domain has an important role in the binding of ss and dsDNA. Together, a functional cross talk between the two domains of HpDprA facilitating the binding and formation of higher order complex with DNA is discussed.

Physical Layer Data Fusion Via Distributed Co-Phasing With General Signal Constellations

This paper studies a pilot-assisted physical layer data fusion technique known as Distributed Co-Phasing (DCP). In this two-phase scheme, the sensors first estimate the channel to the fusion center (FC) using pilots sent by the latter; and then they simultaneously transmit their common data by pre-rotating them by the estimated channel phase, thereby achieving physical layer data fusion. First, by analyzing the symmetric mutual information of the system, it is shown that the use of higher order constellations (HOC) can improve the throughput of DCP compared to the binary signaling considered heretofore. Using an HOC in the DCP setting requires the estimation of the composite DCP channel at the FC for data decoding. To this end, two blind algorithms are proposed: 1) power method, and 2) modified K-means algorithm. The latter algorithm is shown to be computationally efficient and converges significantly faster than the conventional K-means algorithm. Analytical expressions for the probability of error are derived, and it is found that even at moderate to low SNRs, the modified K-means algorithm achieves a probability of error comparable to that achievable with a perfect channel estimate at the FC, while requiring no pilot symbols to be transmitted from the sensor nodes. Also, the problem of signal corruption due to imperfect DCP is investigated, and constellation shaping to minimize the probability of signal corruption is proposed and analyzed. The analysis is validated, and the promising performance of DCP for energy-efficient physical layer data fusion is illustrated, using Monte Carlo simulations.

Solid-state optical absorption from optimally tuned time-dependent range-separated hybrid density functional theory

We present a framework for obtaining reliable solid-state charge and optical excitations and spectra from optimally tuned range-separated hybrid density functional theory. The approach, which is fully couched within the formal framework of generalized Kohn-Sham theory, allows for the accurate prediction of exciton binding energies. We demonstrate our approach through first principles calculations of one- and two-particle excitations in pentacene, a molecular semiconducting crystal, where our work is in excellent agreement with experiments and prior computations. We further show that with one adjustable parameter, set to produce the known band gap, this method accurately predicts band structures and optical spectra of silicon and lithium fluoride, prototypical covalent and ionic solids. Our findings indicate that for a broad range of extended bulk systems, this method may provide a computationally inexpensive alternative to many-body perturbation theory, opening the door to studies of materials of increasing size and complexity.

A Fast and Fault-Tolerant Distributed Algorithm for Near-Optimal TDMA Scheduling in WSNs

The time division multiple access (TDMA) based channel access mechanisms perform better than the contention based channel access mechanisms, in terms of channel utilization, reliability and power consumption, specially for high data rate applications in wireless sensor networks (WSNs). Most of the existing distributed TDMA scheduling techniques can be classified as either static or dynamic. The primary purpose of static TDMA scheduling algorithms is to improve the channel utilization by generating a schedule of smaller length. But, they usually take longer time to schedule, and hence, are not suitable for WSNs, in which the network topology changes dynamically. On the other hand, dynamic TDMA scheduling algorithms generate a schedule quickly, but they are not efficient in terms of generated schedule length. In this paper, we propose a novel scheme for TDMA scheduling in WSNs, which can generate a compact schedule similar to static scheduling algorithms, while its runtime performance can be matched with those of dynamic scheduling algorithms. Furthermore, the proposed distributed TDMA scheduling algorithm has the capability to trade-off schedule length with the time required to generate the schedule. This would allow the developers of WSNs, to tune the performance, as per the requirement of prevalent WSN applications, and the requirement to perform re-scheduling. Finally, the proposed TDMA scheduling is fault-tolerant to packet loss due to erroneous wireless channel. The algorithm has been simulated using the Castalia simulator to compare its performance with those of others in terms of generated schedule length and the time required to generate the TDMA schedule. Simulation results show that the proposed algorithm generates a compact schedule in a very less time.

Co-Exploration of NLA Kernels and Specification of Compute Elements in Distributed Memory CGRAs

Coarse Grained Reconfigurable Architectures (CGRA) are emerging as embedded application processing units in computing platforms for Exascale computing. Such CGRAs are distributed memory multi- core compute elements on a chip that communicate over a Network-on-chip (NoC). Numerical Linear Algebra (NLA) kernels are key to several high performance computing applications. In this paper we propose a systematic methodology to obtain the specification of Compute Elements (CE) for such CGRAs. We analyze block Matrix Multiplication and block LU Decomposition algorithms in the context of a CGRA, and obtain theoretical bounds on communication requirements, and memory sizes for a CE. Support for high performance custom computations common to NLA kernels are met through custom function units (CFUs) in the CEs. We present results to justify the merits of such CFUs.