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At least 397 records · Page 22

Template switching between PNA and RNA oligonucleotides

The origin of the RNA world is not easily understood, as effective prebiotic syntheses of the components of RNA, the beta-ribofuranoside-5'-phosphates, are hard to envisage. Recognition of this difficulty has led to the proposal that other genetic systems, the components of which are more easily formed, may have preceded RNA. This raises the question of how transitions between one genetic system and another could occur. Peptide nucleic acid (PNA) resembles RNA in its ability to form double-helical complexes stabilized by Watson-Crick hydrogen bonding between adenine and thymine and between cytosine and guanine, but has a backbone that is held together by amide rather than by phosphodiester bonds. Oligonucleotides bases on RNA are known to act as templates that catalyse the non-enzymatic synthesis of their complements from activated mononucleotides, we now show that RNA oligonucleotides facilitate the synthesis of complementary PNA strands and vice versa. This suggests that a transition between different genetic systems can occur without loss of information.

NASA Discipline Exobiology↗

Structural Analysis of 23S rRNA Methylating Enzyme Cfr Reveals RNA-Binding Determinants for Methylation Regiospecificity and Antibiotic Resistance

The 23S rRNA methylating enzyme Cfr, found in pathogens including Staphylococcus aureus, Clostridium difficile, Escherichia coli, and Klebsiella pneumoniae, confers resistance to phenicols, lincosamides, oxazolidinones (including linezolid), pleuromutilins, and streptogramins A (the PhLOPS A phenotype). Cfr catalyzes methylation of the C8 position of the A2503 base in 23S rRNA, the recognition site of the above antibiotic classes. Along with the RlmN housekeeping enzyme, Cfr can also promote methylation of the C2 position of the same base. The molecular and structural basis of Cfr’s dual substrate specificity is not known, which hinders our ability to design Cfr-targeting inhibitors necessary to curb PhLOPS A resistance. Here, we present the first crystal structure of Cfr and a detailed analysis of its possible interactions with rRNA. Using structure-guided mutagenesis, mass spectrometry analysis of in cellulo 23S rRNA methylated species, and in cellulo resistance studies, we identify the key amino acids essential for Cfr methylation and multidrug resistance activity. In particular, we found that Cfr’s Q329 residue is important for C8- specific methylation. These data provide a framework for further studies of the biochemistry, structure, and inhibition of this important resistance determinant.

AlphaFold↗

Optical recognition of constructs using hyperspectral imaging and detection (ORCHID)

Abstract Challenges to deep sample imaging have necessitated the development of special techniques such as spatially offset optical spectroscopy to collect signals that have travelled through several layers of tissue. However, these techniques provide only spectral information in one dimension (i.e., depth). Here, we describe a general and practical method, referred to as Optical Recognition of Constructs Using Hyperspectral Imaging and Detection (ORCHID). The sensing strategy integrates (1) the spatial offset detection concept by computationally binning 2D optical data associated with digital offsets based on selected radial pixel distances from the excitation source; (2) hyperspectral imaging using tunable filter; and (3) digital image binding and collation. ORCHID is a versatile modality that is designed to collect optical signals deep inside samples across three spatial (X, Y, Z) as well as spectral dimensions. The ORCHID method is applicable to various optical techniques that exhibit narrow-band structures, from Raman scattering to quantum dot luminescence. Samples containing surface-enhanced Raman scattering (SERS)-active gold nanostar probes and quantum dots embedded in gel were used to show a proof of principle for the ORCHID concept. The resulting hyperspectral data cube is shown to spatially locate target emitting nanoparticle volumes and provide spectral information for in-depth 3D imaging.

47 OTHER INSTRUMENTATION↗

Understanding and Verifying Neural Networks

Deep Neural Networks (DNNs) have gained immense popularity in recent times and have widespread use in applications such as image classification, sentiment analysis, speech recognition and also in safety-critical applications such as autonomous driving. However, they suffer limitations such as lack of explainability and robustness which raise safety and security concerns in their usage. Further, the complex structure and large input spaces of DNNs act as an impediment to thorough verification and testing. The SafeDNN project at the Robust Software Engineering (RSE) group at NASA aims at exploring techniques to ensure that systems that use deep neural networks are safe, robust and interpretable. In this talk, I will be presenting our technique Prophecy that automatically infers formal properties of deep neural network models. The tool extracts patterns based on neuron activations as preconditions that imply certain desirable output properties of the model. I would be highlighting case studies that use Prophecy in obtaining explanations for network decisions, understanding correct and incorrect behavior, providing formal guarantees wrt safety and robustness, and debugging neural network models. We have applied the tool on image classification networks, neural network controllers providing turn advisories in unmanned aircrafts, regression models used for autonomous center-line tracking in aircrafts and neural network object detectors

Deep Neural Networks↗

How μ-opioid receptor recognizes fentanyl

Abstract Roughly half of the drug overdose-related deaths in the United States are related to synthetic opioids represented by fentanyl which is a potent agonist of mu-opioid receptor (mOR). In recent years, X-ray crystal structures of mOR in complex with morphine derivatives have been determined; however, structural basis of mOR activation by fentanyl-like opioids remains lacking. Exploiting the X-ray structure of BU72-bound mOR and several molecular simulation techniques, we elucidated the detailed binding mechanism of fentanyl. Surprisingly, in addition to the salt-bridge binding mode common to morphinan opiates, fentanyl can move deeper and form a stable hydrogen bond with the conserved His297 6.52 , which has been suggested to modulate mOR’s ligand affinity and pH dependence by previous mutagenesis experiments. Intriguingly, this secondary binding mode is only accessible when His297 6.52 adopts a neutral HID tautomer. Alternative binding modes may represent a general mechanism in G protein-coupled receptor-ligand recognition.

60 APPLIED LIFE SCIENCES↗

Architecture-Preserving Provable Repair of Deep Neural Networks

Deep neural networks (DNNs) are becoming increasingly important components of software, and are considered the state-of-the-art solution for a number of problems, such as image recognition. However, DNNs are far from infallible, and incorrect behavior of DNNs can have disastrous real-world consequences. This paper addresses the problem of architecture-preserving V-polytope provable repair of DNNs. A V-polytope defines a convex bounded polytope using its vertex representation. V-polytope provable repair guarantees that the repaired DNN satisfies the given specification on the infinite set of points in the given V-polytope. An architecture-preserving repair only modifies the parameters of the DNN, without modifying its architecture. The repair has the flexibility to modify multiple layers of the DNN, and runs in polynomial time. It supports DNNs with activation functions that have some linear pieces, as well as fully-connected, convolutional, pooling and residual layers. To the best our knowledge, this is the first provable repair approach that has all of these features. We implement our approach in a tool called APRNN. Using MNIST, ImageNet, and ACAS Xu DNNs, we show that it has better efficiency, scalability, and generalization compared to PRDNN and REASSURE, prior provable repair methods that are not architecture preserving.

97 MATHEMATICS AND COMPUTING↗

Arctic and Antarctic Analogs for Planetary Surface Traverses

The proposed paper summarizes the workshop presentations and discusses several of the key findings or lessons including: (1) A recognition that NASA s current approach for long duration planetary surface operations has fundamental differences from any of the operational approaches described by the invited speakers. These approaches drive the crew size and skill mix to accomplish basic objectives and, in turn, drive the logistical pyramid needed to support these operations. NASA will review the operational approaches of the organizations represented to understand the differentiating factors. NASA will then decide if it should alter its current approach to surface exploration. (2) There are potential parallels between key characteristics of the systems used for exploration in these environments, such as heated volume as an analog for pressurized volume or energy usage for various activities. NASA will look at these characteristics to identify which could help with preliminary planning and gather raw data from the presenters to model these characteristics. (3) New technologies are being applied and design approaches are being tailored to take advantage of these technologies on both side. Interactions between these two communities has begun or is expanding to understand how these new technologies are being leveraged: NASA habitation designers are exchanging ideas and approaches with the Antarctic station designers; Antarctic support

Hoffman, Stephen J.↗

NASA Tech Briefs, January 2011

The topics include: 1) Distributed Aerodynamic Sensing and Processing Toolbox; 2) Collaborative Supervised Learning for Sensor Networks; 3) Hazard Detection Software for Lunar Landing; 4) Onboard Nonlinear Engine Sensor and Component Fault Diagnosis and Isolation Scheme; 5) Network-Capable Application Process and Wireless Intelligent Sensors for ISHM; 6) Interface Supports Multiple Broadcast Transceivers for Flight Applications; 7) FPGA Sequencer for Radar Altimeter Applications; 8) Miniature Sapphire Acoustic Resonator - MSAR; 9) Process-Hardened, Multi-Analyte Sensor for Characterizing Rocket Plume Constituents; 10) SAD5 Stereo Correlation Line-Striping in an FPGA; 11) Hybrid Composite Cryogenic Tank Structure; 12) Nanoscale Deformable Optics; 13) Reliability-Based Design Optimization of a Composite Airframe Component; 14) Zinc Oxide Nanowire Interphase for Enhanced Lightweight Polymer Fiber Composites; 15) Plasma Igniter for Reliable Ignition of Combustion in Rocket Engines; 16) Wire Test Grip Fixture; 17) A Sub-Hertz, Low-Frequency Vibration Isolation Platform; 18) Carbon Nanofibers Synthesized on Selective Substrates for Nonvolatile Memory and 3D Electronics; 19) Nanoparticle/Polymer Nanocomposite Bond Coat or Coating; 20) High-Resolution Wind Measurements for Offshore Wind Energy Development; 21) Spring Tire; 22) Marsviewer 2008; 23) Mission Services Evolution Center Message Bus; 24) Major Constituents Analysis for the Vehicle Cabin Atmosphere Monitor; 25) Astronaut Health Participant Summary Application; 26) Adaption of the AMDIS Method to Flight Status on the VCAM Instrument; 27) Natural Language Interface for Safety Certification of Safety-Critical Software; 28) Cryogenic Caging for Science Instrumentation; 29) Wide-Range Neutron Detector for Space Nuclear Applications; 30) In Situ Guided Wave Structural Health Monitoring System; 31) Multiplexed Energy Coupler for Rotating Equipment; 32) Attitude Estimation in Fractionated Spacecraft Cluster Systems; 33) Full Piezoelectric Multilayer-Stacked Hybrid Actuation/Transduction Systems; 34) Active Flow Effectors for Noise and Separation Control; 35) Method and System for Temporal Filtering in Video Compression Systems; 36) Apparatus for Measuring Total Emissivity of Small, Low-Emissivity Samples; 37) Multiple-Zone Diffractive Optic Element for Laser Ranging Applications; 38) Simplified Architecture for Precise Aiming of a Deep-Space Communication Laser Transceiver; 39) Two-Photon-Absorption Scheme for Optical Beam Tracking; 40) High-Sensitivity, Broad-Range Vacuum Gauge Using Nanotubes for Micromachined Cavities; 41) Wide-Field Optic for Autonomous Acquisition of Laser Link; 42) Extracting Zero-Gravity Surface Figure of a Mirror; 43) Modeling Electromagnetic Scattering From Complex Inhomogeneous Objects; 44) Visual Object Recognition and Tracking of Tools; 45) Method for Implementing Optical Phase Adjustment; 46) Visual SLAM Using Variance Grid Maps; 47) Rapid Calculation of Spacecraft Trajectories Using Efficient Taylor Series Integration; 48) Efficient Kriging Algorithms; 49) Predicting Spacecraft Trajectories by the WeavEncke Method; 50) An Augmentation of G-Guidance Algorithms; 51) Comparison of Aircraft Icing Growth Assessment Software; 52) Silicon-Germanium Voltage-Controlled Oscillator at 105 GHz; 53) Estimation of Coriolis Force and Torque Acting on Ares-1; 54) Null Lens Assembly for X-Ray Mirror Segments; and 55) High-Precision Pulse Generator.

Source record↗

Entropy-defect synergy for dual luminescence mechanism in spinel: Time-resolved anti-counterfeiting and fingerprint visualization

Multimodal luminescent materials, while promising for anti-counterfeiting, often lack dynamic time-dependent responses and controllable spatial distribution, limiting their encryption capabilities in the spatiotemporal dimension. Here, this work presents a coordinated control strategy based on entropy and defect engineering, and uses a backpropagation (BP) neural network for material screening to successfully prepare spinel Mg 0.8 (Fe 0.04 Co 0.04 Ni 0.04 Cu 0.04 Zn 0.04 )Cr 2 O 4 (MgA 5 CO) phosphors with time-dependent dynamic luminescence behavior. This phosphor simultaneously activated the d-d transition luminescence (∼618 nm) derived from Co 2+ /Cr 3+ and the defect luminescence (∼398 nm) related to zinc vacancies (V Zn ) in a single-phase solid solution. The phosphor exhibits a time-dependent color evolution from pink to purple under fixed-wavelength excitation, due to the different excited-state dynamics and decay lifetimes associated with the d-d transition and defect luminescence. Structural characterization and spectral analysis confirmed the existence of V Zn and its significant role in defect luminescence process. The fluorescent and dynamic luminescent properties of entropy-based spinel oxide enable its use in advanced anti-counterfeiting applications like fingerprint recognition and color-changing dedicated anti-counterfeiting mark, showing promise in high-end and time-dynamic anti-counterfeiting fields. This research not only developed a new type of fluorescent dynamic anti-counterfeiting material, but also provided a new idea for constructing advanced optical functional materials with multiple luminescence mechanisms.

Defect project↗

NASA total quality management 1989 accomplishments report

NASA and contractor employees achieved many notable improvements in 1989. The highlights of those improvements, described in this seventh annual Accomplishments Report, demonstrate that the people who support NASA's activities are getting more involved in quality and continuous improvement efforts. Their gains solidly support NASA's and this Nation's goal to remain a leader in space exploration and in world-wide market competition, and, when communicated to others through avenues such as this report, foster improvement efforts across government and industry. The principles in practice which led to these process refinements are important cultural elements to any organization's productivity and quality efforts. The categories in this report reflect NASA principles set forth in the 1980's and are more commonly known today as Total Quality Management (TQM): top management leadership and support; strategic planning; focus on the customer; employee training and recognition; employee empowerment and teamwork; measurement and analysis; and quality assurance.

Tai, Betty P.↗

Biophysical and Structural Features of αβT ‐Cell Receptor Mechanosensing: A Paradigmatic Shift in Understanding T‐Cell Activation

ABSTRACT αβT cells protect vertebrates against many diseases, optimizing surveillance using mechanical force to distinguish between pathophysiologic cellular alterations and normal self‐constituents. The multi‐subunit αβT‐cell receptor (TCR) operates outside of thermal equilibrium, harvesting energy via physical forces generated by T‐cell motility and actin‐myosin machinery. When a peptide‐bound major histocompatibility complex molecule (pMHC) on an antigen presenting cell is ligated, the αβTCR on the T cell leverages force to form a catch bond, prolonging bond lifetime, and enhancing antigen discrimination. Under load, the αβTCR undergoes reversible structural transitions involving partial unfolding of its clonotypic immunoglobulin‐like (Ig) domains and coupled rearrangements of associated CD3 subunits and structural elements. We postulate that transitions provide critical energy to initiate the signaling cascade via induction of αβTCR quaternary structural rearrangements, associated membrane perturbations, exposure of CD3 ITAMs to phosphorylation by non‐receptor tyrosine kinases, and phase separation of signaling molecules. Understanding force‐mediated signaling by the αβTCR clarifies long‐standing questions regarding αβTCR antigen recognition, specificity and affinity, providing a basis for continued investigation. Future directions include examining atomistic mechanisms of αβTCR signal initiation, performance quality, tissue compliance adaptability, and T‐cell memory fate. The mechanotransduction paradigm will foster improved rational design of T‐cell based vaccines, CAR‐Ts, and adoptive therapies.

Immunology↗

A robotic system for automation of logistics functions on the Space Station

Spacecraft inventory management is currently performed by the crew and as systems become more complex, increased crew time will be required to perform routine logistics activities. If future spacecraft are to function effectively as research labs and production facilities, the efficient use of crew time as a limited resource for performing mission functions must be employed. The use of automation and robotics technology, such as automated warehouse and materials handling functions, can free the crew from many logistics tasks and provide more efficient use of crew time. Design criteria for a Space Station Automated Logistics Inventory Management System is focused on through the design and demonstration of a mobile two armed terrestrial robot. The system functionally represents a 0 gravity automated inventory management system and the problems associated with operating in such an environment. Features of the system include automated storage and retrieval, item recognition, two armed robotic manipulation, and software control of all inventory item transitions and queries.

Martin, J. C.↗

NASA Tech Briefs, June 2014

Topics include: Real-Time Minimization of Tracking Error for Aircraft Systems; Detecting an Extreme Minority Class in Hyperspectral Data Using Machine Learning; KSC Spaceport Weather Data Archive; Visualizing Acquisition, Processing, and Network Statistics Through Database Queries; Simulating Data Flow via Multiple Secure Connections; Systems and Services for Near-Real-Time Web Access to NPP Data; CCSDS Telemetry Decoder VHDL Core; Thermal Response of a High-Power Switch to Short Pulses; Solar Panel and System Design to Reduce Heating and Optimize Corridors for Lower-Risk Planetary Aerobraking; Low-Cost, Very Large Diamond-Turned Metal Mirror; Very-High-Load-Capacity Air Bearing Spindle for Large Diamond Turning Machines; Elevated-Temperature, Highly Emissive Coating for Energy Dissipation of Large Surfaces; Catalyst for Treatment and Control of Post-Combustion Emissions; Thermally Activated Crack Healing Mechanism for Metallic Materials; Subsurface Imaging of Nanocomposites; Self-Healing Glass Sealants for Solid Oxide Fuel Cells and Electrolyzer Cells; Micromachined Thermopile Arrays with Novel Thermo - electric Materials; Low-Cost, High-Performance MMOD Shielding; Head-Mounted Display Latency Measurement Rig; Workspace-Safe Operation of a Force- or Impedance-Controlled Robot; Cryogenic Mixing Pump with No Moving Parts; Seal Design Feature for Redundancy Verification; Dexterous Humanoid Robot; Tethered Vehicle Control and Tracking System; Lunar Organic Waste Reformer; Digital Laser Frequency Stabilization via Cavity Locking Employing Low-Frequency Direct Modulation; Deep UV Discharge Lamps in Capillary Quartz Tubes with Light Output Coupled to an Optical Fiber; Speech Acquisition and Automatic Speech Recognition for Integrated Spacesuit Audio Systems, Version II; Advanced Sensor Technology for Algal Biotechnology; High-Speed Spectral Mapper; "Ascent - Commemorating Shuttle" - A NASA Film and Multimedia Project DVD; High-Pressure, Reduced-Kinetics Mechanism for N-Hexadecane Oxidation; Method of Error Floor Mitigation in Low-Density Parity-Check Codes; X-Ray Flaw Size Parameter for POD Studies; Large Eddy Simulation Composition Equations for Two-Phase Fully Multicomponent Turbulent Flows; Scheduling Targeted and Mapping Observations with State, Resource, and Timing Constraints;

Source record↗

Unique structural solution from a V H 3-30 antibody targeting the hemagglutinin stem of influenza A viruses

Broadly neutralizing antibodies (bnAbs) targeting conserved influenza A virus (IAV) hemagglutinin (HA) epitopes can provide valuable information for accelerating universal vaccine designs. Here, we report structural details for heterosubtypic recognition of HA from circulating and emerging IAVs by the human antibody 3I14. Somatic hypermutations play a critical role in shaping the HCDR3, which alone and uniquely among V H 3-30 derived antibodies, forms contacts with five sub-pockets within the HA-stem hydrophobic groove. 3I14 light-chain interactions are also key for binding HA and contribute a large buried surface area spanning two HA protomers. Comparison of 3I14 to bnAbs from several defined classes provide insights to the bias selection of V H 3-30 antibodies and reveals that 3I14 represents a novel structural solution within the V H 3-30 repertoire. The structures reported here improve our understanding of cross-group heterosubtypic binding activity, providing the basis for advancing immunogen designs aimed at eliciting a broadly protective response to IAV.

59 BASIC BIOLOGICAL SCIENCES↗

Supplier Research & Analysis Approach

"The implementation of NASA GSFC's portfolio of mission projects relies upon inter-connected, multi-tiered supply chains of organizations operating under direct and indirect contracts and other agreements throughout the U.S. and around the world. These supply chains are subject to an inter-related array of technical/production, business, market and security risks that are amplified by the ongoing globalization of industry and technology and which can disrupt or threaten the production and delivery of products and services when needed and in conformance with requirements. In recognition of such risks and associated challenges, GSFC's SMA directorate launched an innovative Supplier Research and Analysis (SRA) capability three years ago to gain greater insight into the operating environment, performance, capabilities and viability of current and prospective suppliers for GSFC projects and proposals. The capability uses business intelligence techniques and primarily open source information resources as part of a cost-effective, non-intrusive methodology to produce several types of research and analysis reports. The reports are based on a holistic analytical framework encompassing key technical/production, business enterprise management, market and security factors, and feature in-depth information, summary information profiles, SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis, and candidate risk concerns in order to pro-actively support SMA and project management needs. The SRA capability, which is designed to complement and support ongoing SMA/project management activities and practices, has produced over 95 reports since its start-up in early 2015. This presentation addresses the approach, methodology and performance of the Supplier Research and Analysis (SRA) capability and its value in assuring the success of NASA mission projects. In doing so, the presentation provides lessons-learned, best practices, case examples and address how it fits into the development of an enterprise-level Supply Chain Risk Management capability."

supplier research and analysis↗

Supply Chain Research and Analysis for Space Systems

The implementation of NASA GSFC's portfolio of mission projects relies upon inter-connected, multi-tiered supply chains of organizations operating under direct and indirect contracts and other agreements throughout the U.S. and around the world. These supply chains are subject to an inter-related array of technical/production, business, market and security risks that are amplified by the ongoing globalization of industry and technology and which can disrupt or threaten the production and delivery of products and services when needed and in conformance with requirements. In recognition of such risks and associated challenges, GSFC's SMA directorate launched an innovative Supply Chain Research and Analysis capability three years ago to gain greater insight into the operating environment, performance, capabilities and viability of current and prospective suppliers for GSFC projects and proposals. The capability uses business intelligence techniques and primarily open source information resources as part of a cost-effective, non-intrusive methodology to produce several types of research and analysis reports. The reports are based on a holistic analytical framework encompassing key technical/production, business enterprise management, market and security factors, and feature in-depth information, summary information profiles, SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis, and candidate risk concerns in order to pro-actively support SMA and project management needs. The SRA capability, which is designed to complement and support ongoing SMA/project management activities and practices, has produced over 105 reports since its start-up in early 2015.This presentation addresses the approach, methodology and performance of the Supply Chain Research and Analysiscapability and its value in assuring the success of NASA mission projects. In doing so, the presentation provides lessons-learned, best practices, case examples and address how it fits into the development of an enterprise-level Supply Chain Risk Management capability.

supply chain risk management↗

Towards Probablistic Assessment of Hypobaric Decompression Sickness Treatment

INTRODUCTION: Pressure, oxygen (O2), and time are the pillars to effective treatment of decompression sickness (DCS). The NASA DCS Treatment Model links a decrease in computed bubble volume to the resolution of a symptom. The decrease in volume is realized in two stages: a) during the Boyle's Law compression and b) during subsequent dissolution of the gas phase by the O2 window. METHODS: The cumulative distribution of 154 symptoms that resolved during repressurization was described with a log-logistic density function of pressure difference (deltaP as psid) associated with symptom resolution and two other explanatory variables. The 154 symptoms originated from 119 cases of DCS during 969 exposures in 47 different altitude tests. RESULTS: The probability of symptom resolution [P(symptom resolution)] = 1 / (1+exp(- (ln(deltaP) - 1.682 + 1.089×AMB - 0.00395×SYMPTOM TIME) / 0.633)), where AMB is 1 when the subject ambulated as part of the altitude exposure or else 0 and SYMPTOM TIME is the elapsed time in min from start of the altitude exposure to recognition of a DCS symptom. The P(symptom resolution) was estimated from computed deltaP from the Tissue Bubble Dynamics Model based on the "effective" Boyle's Law change: P2 - P1 (deltaP, psid) = P1×V1/V2 - P1, where V1 is the computed volume of a spherical bubble in a unit volume of tissue at low pressure P1 and V2 is computed volume after a change to a higher pressure P2. V2 continues to decrease through time at P2, at a faster rate if 100% ground level O2 was breathed. The computed deltaP is the effective treatment pressure at any point in time as if the entire deltaP was just from Boyle's Law compression. DISCUSSION: Given the low probability of DCS during extravehicular activity and the prompt treatment of a symptom with options through the model it is likely that the symptom and gas phase will resolve with minimum resources and minimal impact on astronaut health, safety, and productivity.

Conkin, J.↗

A distinct RNA recognition mechanism governs Np 4 decapping by RppH

Dinucleoside tetraphosphates, often described as alarmones because their cellular concentration increases in response to stress, have recently been shown to function in bacteria as precursors to nucleoside tetraphosphate (Np 4 ) RNA caps. Removal of this cap is critical for initiating 5' end-dependent degradation of those RNAs, potentially affecting bacterial adaptability to stress; however, the predominant Np 4 decapping enzyme in proteobacteria, ApaH, is inactivated by the very conditions of disulfide stress that enable Np 4 -capped RNAs to accumulate to high levels. Here, in this study, we show that, in Escherichia coli cells experiencing such stress, the RNA pyrophosphohydrolase RppH assumes a leading role in decapping those transcripts, preferring them as substrates over their triphosphorylated and diphosphorylated counterparts. Unexpectedly, this enzyme recognizes Np 4 -capped 5' ends by a mechanism distinct from the one it uses to recognize other 5' termini, resulting in a one-nucleotide shift in substrate specificity. The unique manner in which capped substrates of this kind bind to the active site of RppH positions the δ-phosphate, rather than the β-phosphate, for hydrolytic attack, generating triphosphorylated RNA as the primary product of decapping. Consequently, a second RppH-catalyzed deprotection step is required to produce the monophosphorylated 5' terminus needed to stimulate rapid RNA decay. The unconventional manner in which RppH recognizes Np 4 -capped 5' ends and its differential impact on the rates at which such termini are deprotected as a prelude to RNA degradation could have major consequences for reprogramming gene expression during disulfide stress.

59 BASIC BIOLOGICAL SCIENCES↗