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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 91 records · Page 5
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Structures of RNA phosphotransferase Tpt1 reveal distinct binding modes for an RNA 2′-PO 4 splice junction versus a 5′-PO 4 mononucleotide

Tpt1 is a widely distributed enzyme that removes an internal RNA 2′-phosphate by transfer to NAD + , via a two-step reaction in which: (i) the RNA 2′-PO 4 attacks NAD + to form an RNA-2′-phospho-(ADP-ribose) intermediate and expel nicotinamide; and (ii) the ADP-ribose O2″ attacks the RNA 2′-phosphodiester to form 2′-OH RNA and ADP-ribose-1″,2″-cyclic phosphate products. Tpt1 can also execute a single-step ADP-ribosyltransferase reaction at a 5′-monophosphate nucleic acid terminus that installs a 5′-phospho-ADP-ribose cap structure. Here we present crystal structures of Tpt1 bound to an RNA containing an internal 2′-PO 4 mark (the substrate for the canonical Tpt1 pathway) and in a complex with 5′-AMP. We find that Tpt1 has distinct binding modes, whereby the RNA 2′-PO 4 and the AMP 5′-PO 4 are engaged by the same set of active site amino acids, but the 2′-PO 4 nucleoside and the 5′-nucleoside occupy different sites on the enzyme.

Biochemistry & Molecular Biology↗
NASA NTRS1980

Cable-splice detector

Detector has possible uses in aerial cable-car systems, equipment handling in mines, boreholes, and undersea operations, and other applications where moving steel cable must be measured, monitored, or controlled. Detector consists of Hall-effect magnetic sensor located close to cable. Magnetic markings on cable are converted to electrical signals. Signals are filtered, amplified, and can actuate alarm.

Lee, R. D.↗
NASA NTRS1986

Strength of graphite/epoxy bolted wing-skin splice specimens subjected to outdoor exposure under constant load and yearly fatigue loading

The results of an experimental study to provide long-term durability data on detailed full-scale graphite/epoxy wing-skin joint designs under environmental exposure and cyclic loading associated with commercial transport aircraft are reported. The specimens consisted of a single-row bolt configuration fabricated from T300/5208 and a double-row bolt configuration fabricated from T300/5209. The unpainted specimens were exposed to the outdoor environment under a sustained tensile load, and at yearly intervals, they were subjected to fatigue loading. Experimental results showed a slight reduction in residual tensile strength for both graphite/epoxy joints under the exposure times and fatigue loadings reported. A 7.5-percent decrease in residual strength was observed for the T300/5208 single-row joint after 5 years exposure and two lifetimes of fatigue loading. A 5.3-percent decrease in residual strength was observed for the T300/5209 double-row joint after 7 years exposure and 2.8 lifetimes of fatigue loading. The 5208 epoxy material was more susceptible to degradation by ultraviolet radiation than the 5209 epoxy material.

Wichorek, G. R.↗
NASA NTRS1992

Metal-To-Composite Shaft Splice

Strong, lightweight shaft formed without awkward bolted flanges. Composite and metal portions of shaft joined by winding composite portion onto outer surface and into spiral groove of metal portion. Groove and composite material constitute structural bond. If opposing spiral grooves used, then two shafts locked together against torques in both directions.

Kish, Jules↗
NASA NTRS1993

Modeling study on the cleavage step of the self-splicing reaction in group I introns

A three-dimensional model of the Tetrahymena thermophila group I intron is used to further explore the catalytic mechanism of the transphosphorylation reaction of the cleavage step. Based on the coordinates of the catalytic core model proposed by Michel and Westhof (Michel, F., Westhof, E. J. Mol. Biol. 216, 585-610 (1990)), we first converted their ligation step model into a model of the cleavage step by the substitution of several bases and the removal of helix P9. Next, an attempt to place a trigonal bipyramidal transition state model in the active site revealed that this modified model for the cleavage step could not accommodate the transition state due to insufficient space. A lowering of P1 helix relative to surrounding helices provided the additional space required. Simultaneously, it provided a better starting geometry to model the molecular contacts proposed by Pyle et al. (Pyle, A. M., Murphy, F. L., Cech, T. R. Nature 358, 123-128. (1992)), based on mutational studies involving the J8/7 segment. Two hydrated Mg2+ complexes were placed in the active site of the ribozyme model, using the crystal structure of the functionally similar Klenow fragment (Beese, L.S., Steitz, T.A. EMBO J. 10, 25-33 (1991)) as a guide. The presence of two metal ions in the active site of the intron differs from previous models, which incorporate one metal ion in the catalytic site to fulfill the postulated roles of Mg2+ in catalysis. The reaction profile is simulated based on a trigonal bipyramidal transition state, and the role of the hydrated Mg2+ complexes in catalysis is further explored using molecular orbital calculations.

NASA Discipline Exobiology↗
NASA NTRS2017

The Profile Envision and Splicing Tool (PRESTO): Developing an Atmospheric Wind Analysis Tool for Space Launch Vehicles Using Python

Launch vehicle programs require vertically complete atmospheric profiles. Many systems at the ER to make the necessary measurements, but all have different EVR, vertical coverage, and temporal coverage. MSFC Natural Environments Branch developed a tool to create a vertically complete profile from multiple inputs using Python. Forward work: Finish Formal Testing Acceptance Testing, End-to-End Testing. Formal Release

Orcutt, John M.↗
DOE OSTI2022

A high-resolution single-molecule sequencing-based Arabidopsis transcriptome using novel methods of Iso-seq analysis

Accurate and comprehensive annotation of transcript sequences is essential for transcript quantification and differential gene and transcript expression analysis. Single-molecule long-read sequencing technologies provide improved integrity of transcript structures including alternative splicing, and transcription start and polyadenylation sites. However, accuracy is significantly affected by sequencing errors, mRNA degradation, or incomplete cDNA synthesis. We present a new and comprehensive Arabidopsis thaliana Reference Transcript Dataset 3 (AtRTD3). AtRTD3 contains over 169,000 transcripts—twice that of the best current Arabidopsis transcriptome and including over 1500 novel genes. Seventy-eight percent of transcripts are from Iso-seq with accurately defined splice junctions and transcription start and end sites. We develop novel methods to determine splice junctions and transcription start and end sites accurately. Mismatch profiles around splice junctions provide a powerful feature to distinguish correct splice junctions and remove false splice junctions. Stratified approaches identify high-confidence transcription start and end sites and remove fragmentary transcripts due to degradation. AtRTD3 is a major improvement over existing transcriptomes as demonstrated by analysis of an Arabidopsis cold response RNA-seq time-series. AtRTD3 provides higher resolution of transcript expression profiling and identifies cold-induced differential transcription start and polyadenylation site usage. AtRTD3 is the most comprehensive Arabidopsis transcriptome currently. It improves the precision of differential gene and transcript expression, differential alternative splicing, and transcription start/end site usage analysis from RNA-seq data. The novel methods for identifying accurate splice junctions and transcription start/end sites are widely applicable and will improve single-molecule sequencing analysis from any species.

transcription start and end sites↗
DOE OSTI2020

Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts

The decoy exon model has been proposed to regulate a subset of intron retention (IR) events involving predominantly larger introns (>1 kb). Splicing reporter studies have shown that decoy splice sites are essential for activity, suggesting that decoys act by engaging intron-terminal splice sites and competing with cross-intron interactions required for intron excision. The decoy model predicts that antisense oligonucleotides may be able to block decoy splice sites in endogenous pre-mRNA, thereby reducing IR and increasing productive gene expression. Indeed, we now demonstrate that targeting a decoy 5' splice site in the O-GlcNAc transferase (OGT) gene reduced IR from ~80% to ~20% in primary human erythroblasts, accompanied by increases in spliced OGT RNA and OGT protein expression. The remaining OGT IR was refractory to antisense treatment and might be mediated by independent mechanism(s). In contrast, other retained introns were strongly dependent on decoy function, since antisense targeting of decoy 5' splice sites greatly reduced (SNRNP70) or nearly eliminated (SF3B1) IR in two widely expressed splicing factors, and also greatly reduced IR in transcripts encoding the erythroid-specific structural protein, α-spectrin (SPTA1). These results show that modulating decoy exon function can dramatically alter IR and suggest that dynamic regulation of decoy exons could be a mechanism to fine-tune gene expression post-transcriptionally in many cell types.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRSDate not supplied

NASA and Blue Origin Collaborative Assessment of Precision Landing Algorithms and Computing

NASA’s Safe and Precise Landing Integrated Capabilities Evolution (SPLICE) project is developing sensor, algorithm, and compute technologies for precision landing and hazard avoidance. These technologies are being tested as an integrated Precision Landing and Hazard Avoidance (PL&HA) system on Blue Origin’s New Shephard suborbital vehicle. A key goal for the computing element of this technology development is to characterize the performance of the SPLICE software workloads on the project’s Descent and Landing Computer (DLC). The DLC is a multi-core processor designed as a surrogate for NASA’s High-Performance Space Computer (HPSC). Measurements of the SPLICE workload performance on the DLC provides NASA insight on how PL&HA capabilities will perform on the HPSC, and guidance on how the SPLICE algorithms can be implemented to best utilize the DLC platform. This insight can also be used to derive requirements to guide trade studies on candidate computing architectures, for use on platforms like Blue Moon. NASA and Blue Origin are collaborating under an agreement to pursue this mutual benefit. Performance metrics collected are based on measurement of common compute resources such as percentage used of memory bandwidth, I/O utilization, interrupt latency, and kernel vs. user space code residency. Where possible existing performance counters and metrics that are part of the operating system kernel are used. As the design has a significant FPGA component, performance counters are identified and instantiated in the fabric to measure DMA performance and interface metrics. Collection of metrics is performed on the DLC with a representative workload that simulates a full landing cycle of the Blue Origin New Shepard vehicle. Consideration is given to the other compute implementations and whether they can run SPLICE algorithms at the same rate and with the same latency as the DLC. One option being considered is the use of a RISC-V soft core instantiated in a radiation resilient FPGA fabric such as the Xilinx KU60. Select algorithms from the SPLICE code will be run for comparison with the DLC. This paper describes how the DLC is instrumented to collect performance measurements of the SPLICE workloads, preliminary results from these measurements, and their implications on SPLICE algorithm implementation. The results of experimentation to derive candidate requirements for architecture trades on a PL&HA computing system are also presented.

computer performance↗
NASA NTRS1979

New Life For The Cable Cars

NASA-Arnes' major recommendations involved ways of extending cable life in the interests of safety and economy. Other recommendations included redesign of the cablegripping device, substitution of modern braking mechanisms, improvements in cable pulleys and other components, and new inspection and repair procedures. Ames followed up by designing and installing new equipment to lengthen cable life, which averages only about two months. These cables-four of them for four different car routes--are endless belts, like ski lift cables, running from the downtown car barn to the end of each line. When a cable is installed, the loop is closed by splicing the ends together in a 72-footlong splice. The splice is the weakest part of the cable and a source of problems. When the car operator applies his grip while over a splice, the resulting friction sometimes causes the splice to "unbraid" and fail; this means shutting down the line until the splice can be repaired. Even when unbraiding does not occur, gripping a splice shortens cable life by friction wear. Worn cables are a safety hazard and must be replaced, which is expensive at $1.60 a foot for 10,000 to 20,000 feet of cable.

Source record↗
NASA NTRS2015

Development of a Climatology of Vertically Complete Wind Profiles from Doppler Radar Wind Profiler Systems

This paper describes in detail the QC and splicing methodology for KSC's 50- and 915-MHz DRWP measurements that generates an extensive archive of vertically complete profiles from 0.20-18.45 km. The concurrent POR from each archive extends from April 2000 to December 2009. MSFC NE applies separate but similar QC processes to each of the 50- and 915-MHz DRWP archives. DRWP literature and data examination provide the basis for developing and applying the automated and manual QC processes on both archives. Depending on the month, the QC'ed 50- and 915-MHz DRWP archives retain 52-65% and 16-30% of the possible data, respectively. The 50- and 915-MHz DRWP QC archives retain 84-91% and 85-95%, respectively, of all the available data provided that data exist in the non- QC'ed archives. Next, MSFC NE applies an algorithm to splice concurrent measurements from both DRWP sources. Last, MSFC NE generates a composite profile from the (up to) five available spliced profiles to effectively characterize boundary layer winds and to utilize all possible 915-MHz DRWP measurements at each timestamp. During a given month, roughly 23,000-32,000 complete profiles exist from 0.25-18.45 km from the composite profiles' archive, and approximately 5,000- 27,000 complete profiles exist from an archive utilizing an individual 915-MHz DRWP. One can extract a variety of profile combinations (pairs, triplets, etc.) from this sample for a given application. The sample of vertically complete DRWP wind measurements not only gives launch vehicle customers greater confidence in loads and trajectory assessments versus using balloon output, but also provides flexibility to simulate different DOL situations across applicable altitudes. In addition to increasing sample size and providing more flexibility for DOL simulations in the vehicle design phase, the spliced DRWP database provides any upcoming launch vehicle program with the capability to utilize DRWP profiles on DOL to compute vehicle steering commands, provided the program applies the procedures that this report describes to new DRWP data on DOL. Decker et al. (2015) details how SLS is proposing to use DRWP data and splicing techniques on DOL. Although automation could enhance the current DOL 50-MHz DRWP QC process and could streamline any future DOL 915-MHz DRWP QC and splicing process, the DOL community would still require manual intervention to ensure that the vehicle only uses valid profiles. If a program desires to use high spatial resolution profiles, then the algorithm could randomly add high-frequency components to the DRWP profiles. The spliced DRWP database provides lots of flexibility in how one performs DOL simulations, and the algorithms that this report provides will assist the aerospace and atmospheric communities that are interested in utilizing the DRWP.

Barbre, Robert E., Jr.↗
DOE OSTI2020

The integral spliceosomal component CWC15 is required for development in Arabidopsis

Efficient mRNA splicing is a prerequisite for protein biosynthesis and the eukaryotic splicing machinery is evolutionarily conserved among species of various phyla. At its catalytic core resides the activated splicing complex Bact consisting of the three small nuclear ribonucleoprotein complexes (snRNPs) U2, U5 and U6 and the so-called NineTeen complex (NTC) which is important for spliceosomal activation. CWC15 is an integral part of the NTC in humans and it is associated with the NTC in other species. Here we show the ubiquitous expression and developmental importance of the Arabidopsis ortholog of yeast CWC15. CWC15 associates with core components of the Arabidopsis NTC and its loss leads to inefficient splicing. Consistent with the central role of CWC15 in RNA splicing, cwc15 mutants are embryo lethal and additionally display strong defects in the female haploid phase. Interestingly, the haploid male gametophyte or pollen in Arabidopsis, on the other hand, can cope without functional CWC15, suggesting that developing pollen might be more tolerant to CWC15-mediated defects in splicing than either embryo or female gametophyte.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Biallelic variants in SNUPN cause a limb girdle muscular dystrophy with myofibrillar-like features

Abstract Alterations in RNA-splicing are a molecular hallmark of several neurological diseases, including muscular dystrophies, where mutations in genes involved in RNA metabolism or characterized by alterations in RNA splicing have been described. Here, we present five patients from two unrelated families with a limb-girdle muscular dystrophy (LGMD) phenotype carrying a biallelic variant in SNUPN gene. Snurportin-1, the protein encoded by SNUPN, plays an important role in the nuclear transport of small nuclear ribonucleoproteins (snRNPs), essential components of the spliceosome. We combine deep phenotyping, including clinical features, histopathology and muscle MRI, with functional studies in patient-derived cells and muscle biopsies to demonstrate that variants in SNUPN are the cause of a new type of LGMD according to current definition. Moreover, an in vivo model in Drosophila melanogaster further supports the relevance of Snurportin-1 in muscle. SNUPN patients show a similar phenotype characterized by proximal weakness starting in childhood, restrictive respiratory dysfunction and prominent contractures, although inter-individual variability in terms of severity even in individuals from the same family was found. Muscle biopsy showed myofibrillar-like features consisting of myotilin deposits and Z-disc disorganization. MRI showed predominant impairment of paravertebral, vasti, sartorius, gracilis, peroneal and medial gastrocnemius muscles. Conservation and structural analyses of Snurportin-1 p.Ile309Ser variant suggest an effect in nuclear-cytosol snRNP trafficking. In patient-derived fibroblasts and muscle, cytoplasmic accumulation of snRNP components is observed, while total expression of Snurportin-1 and snRNPs remains unchanged, which demonstrates a functional impact of SNUPN variant in snRNP metabolism. Furthermore, RNA-splicing analysis in patients’ muscle showed widespread splicing deregulation, in particular in genes relevant for muscle development and splicing factors that participate in the early steps of spliceosome assembly. In conclusion, we report that SNUPN variants are a new cause of limb girdle muscular dystrophy with specific clinical, histopathological and imaging features, supporting SNUPN as a new gene to be included in genetic testing of myopathies. These results further support the relevance of splicing-related proteins in muscle disorders.

Iruzubieta, Pablo (ORCID:0000000303316222)↗