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At least 19 records

Unraveling the mechanism of recognition of the 3’ splice site of the adenovirus major late promoter intron by the alternative splicing factor PUF60

Pre-mRNA splicing is critical for achieving required amounts of a transcript at a given time and for regulating production of encoded protein. A given pre-mRNA may be spliced in many ways, or not at all, giving rise to multiple gene products. Numerous splicing factors are recruited to pre-mRNA splice sites to ensure proper splicing. One such factor, the 60 kDa poly(U)-binding splicing factor (PUF60), is recruited to sites that are not always spliced, but rather function as alternative splice sites. In this study, we characterized the interaction of PUF60 with a splice site from the adenovirus major late promoter (the AdML 3' splice site, AdML3’). We found that the PUF60–AdML3’ dissociation constants are in the micromolar range, with the binding affinity predominantly provided by PUF60’s two central RNA recognition motifs (RRMs). A 1.95 Å crystal structure of the two PUF60 RRMs in complex with AdML3’ revealed a dimeric organization placing two stretches of nucleic acid tracts in opposing directionalities, which can cause looping of nucleic acid and explain how PUF60 affects pre-mRNA geometry to effect splicing. Solution characterization of this complex by light-scattering and UV/Vis spectroscopy suggested a potential 2:1 (PUF60 2 :AdML3’) stoichiometry, consistent with the crystal structure. This work defines the sequence specificity of the alternative splicing factor PUF60 at the pre-mRNA 3’ splice site. Our observations suggest that control of pre-mRNA directionality is important in the early stage of spliceosome assembly, and advance our understanding of the molecular mechanism by which alternative and constitutive splicing factors differentiate among 3’ splice sites.

59 BASIC BIOLOGICAL SCIENCES↗

Stress-Induced Changes in Alternative Splicing Landscape in Rice: Functional Significance of Splice Isoforms in Stress Tolerance

Improvements in yield and quality of rice are crucial for global food security. However, global rice production is substantially hindered by various biotic and abiotic stresses. Making further improvements in rice yield is a major challenge to the rice research community, which can be accomplished through developing abiotic stress-resilient rice varieties and engineering durable agrochemical-independent pathogen resistance in high-yielding elite rice varieties. This, in turn, needs increased understanding of the mechanisms by which stresses affect rice growth and development. Alternative splicing (AS), a post-transcriptional gene regulatory mechanism, allows rapid changes in the transcriptome and can generate novel regulatory mechanisms to confer plasticity to plant growth and development. Mounting evidence indicates that AS has a prominent role in regulating rice growth and development under stress conditions. Several regulatory and structural genes and splicing factors of rice undergo different types of stress-induced AS events, and the functional significance of some of them in stress tolerance has been defined. Both rice and its pathogens use this complex regulatory mechanism to devise strategies against each other. This review covers the current understanding and evidence for the involvement of AS in biotic and abiotic stress-responsive genes, and its relevance to rice growth and development. Furthermore, we discuss implications of AS for the virulence of different rice pathogens and highlight the areas of further research and potential future avenues to develop climate-smart and disease-resistant rice varieties.

59 BASIC BIOLOGICAL SCIENCES↗

Survey of Aging and Monitoring Concerns for Cables and Splices Due to Cable Repair and Replacement

The purpose of this report is to survey aging and monitoring concerns for electrical cable splices in nuclear power plants (NPPs) in long term operation. As portions of existing electrical cable runs in nuclear are replaced over time due to localized events, the total number of splices in NPPs is expected to increase. Relative to cables, the body of knowledge regarding aging of splices and splices in combination with aging cables in nuclear service environments in long-term operations is low. A few reports have considered the aging of cable system components other than cables (Jacobus 1990; Nelson 1998; Villaran and Lofaro 2002), but the nuclear industry has two decades of operating experience since these were published to further enlighten this issue. Herein we discuss electrical cables and splices commonly found in U.S. nuclear power plants, their qualification in safety-related application, and methods for monitoring their health condition. Common environmental stresses that can give rise to cable and splice failure are discussed. The Nuclear Regulatory Commission (NRC) Licensee Event Reports (LER) database was used to identify documented issues of cable and splice failure. The trend in the resultant data over time is considered to see if failures are increasing as plants age. Observations and conclusions of this work include: 1. Cables and splices are highly reliable components. Occurrence rates for events of interest were low and nearly constant over the last 20 years. 2. Common-cause failure for evaluated cable events of interest was observed to primarily be associated with loose connections, which may manifest associated with workmanship issues, thermal cycling, and/or vibration. 3. Replacement of cables is more common than repair, leading to an increase in proportion of new generation cables in the plant over time. 4. Splices on degraded cables have been observed to be problematic. Due to aging NPP infrastructure, including electrical cables, it is expected that such issues will continue to increase. 5. Condition monitoring approaches, while shown to be fruitful for cables, have been shown to be insensitive to degradation of splice sleeves, which are critical to the continued performance of splices. Additional condition monitoring (CM) work is needed to evaluate methods which are sensitive to the degradation of splice components. 6. Extended Material Degradation Assessment (EMDA) knowledge gaps for electrical cables (Bernstein et al. 2014) have not been investigated for splices but may represent similar concerns such as for the accelerated aging process historically used in environmental qualification.

42 ENGINEERING↗

A Deep Exon Cryptic Splice Site Promotes Aberrant Intron Retention in a Von Willebrand Disease Patient

A translationally silent single nucleotide mutation in exon 44 (E44) of the von Willebrand factor (VWF) gene is associated with inefficient removal of intron 44 in a von Willebrand disease (VWD) patient. This intron retention (IR) event was previously attributed to reordered E44 secondary structure that sequesters the normal splice donor site. We propose an alternative mechanism: the mutation introduces a cryptic splice donor site that interferes with the function of the annotated site to favor IR. We evaluated both models using minigene splicing reporters engineered to vary in secondary structure and/or cryptic splice site content. Analysis of splicing efficiency in transfected K562 cells suggested that the mutation-generated cryptic splice site in E44 was sufficient to induce substantial IR. Mutations predicted to vary secondary structure at the annotated site also had modest effects on IR and shifted the balance of residual splicing between the cryptic site and annotated site, supporting competition among the sites. Further studies demonstrated that introduction of cryptic splice donor motifs at other positions in E44 did not promote IR, indicating that interference with the annotated site is context dependent. We conclude that mutant deep exon splice sites can interfere with proper splicing by inducing IR.

60 APPLIED LIFE SCIENCES↗

RNA Splicing Events in Circulation Distinguish Individuals With and Without New-onset Type 1 Diabetes

Context: Alterations in RNA splicing may influence protein isoform diversity that contributes to or reflects the pathophysiology of certain diseases. Whereas specific RNA splicing events in pancreatic islets have been investigated in models of inflammation in vitro, how RNA splicing in the circulation correlates with or is reflective of type 1 diabetes (T1D) disease pathophysiology in humans remains unexplored. Objective: To use machine learning to investigate if alternative RNA splicing events differ between individuals with and without new-onset T1D and to determine if these splicing events provide insight into T1D pathophysiology. Methods: RNA deep sequencing was performed on whole blood samples from 2 independent cohorts: a training cohort consisting of 12 individuals with new-onset T1D and 12 age- and sex-matched nondiabetic controls and a validation cohort of the same size and demographics. Machine learning analysis was used to identify specific isoforms that could distinguish individuals with T1D from controls. Results: Distinct patterns of RNA splicing differentiated participants with T1D from unaffected controls. Notably, certain splicing events, particularly involving retained introns, showed significant association with T1D. Machine learning analysis using these splicing events as features from the training cohort demonstrated high accuracy in distinguishing between T1D subjects and controls in the validation cohort. Gene Ontology pathway enrichment analysis of the retained intron category showed evidence for a systemic viral response in T1D subjects. Conclusion: Alternative RNA splicing events in whole blood are significantly enriched in individuals with new-onset T1D and can effectively distinguish these individuals from unaffected controls. Further, our findings also suggest that RNA splicing profiles offer the potential to provide insights into disease pathogenesis.

60 APPLIED LIFE SCIENCES↗

Conserved Function of RNA Binding Motif Protein48 (RBM48) and Armadillo Repeat Containing 7 (ARMC7) in Maize and Human U12 Splicing

Splicing of pre-mRNA is fundamental for genes containing introns. Emerging data points to a deeply conserved role of this process in eukaryotic cell differentiation and proliferation. The vast majority of introns termed U2-type introns are spliced by a major spliceosome; however, there also exist rare and more conserved U12-type introns that are spliced by a minor spliceosome. Mutations that disrupt U12 splicing inhibit cell differentiation in both maize endosperm and human blood cells. However, the mechanism underlying this process is not well understood. The maize RNA Binding Motif Protein 48 (RBM48) plays an essential role as a U12 splicing factor and is required for proper maize kernel development. Using human cell lines, CRISPR-Cas9 knockdown of RBM48 demonstrated a conserved function in human U12 intron splicing. RBM48 and Armadillo Repeat Containing 7 (ARMC7) protein interact in both maize and human as part of the activated minor spliceosome. Here we show RBM48 and ARMC7 co-localization in the nucleus of human cell lines. Vertebrate ARMC7 is normally localized in the cytosol, whereas RBM48 is found in the nucleus. Our data suggests that the interaction plays a role in regulating ARMC7 localization or the efficiency of U12 intron splicing. We also performed comprehensive transcriptome profiling and identified a common subset of conserved Minor Intron Containing Genes (MIGs) impacted in both human and maize RBM48 knockout mutants. Of importance, the vast majority of these MIGs are associated with developmental defects in both plants and animals. This suggests that aberrant splicing of these targets has a high likelihood of mediating abnormal cell phenotypes. These data support evolutionarily conserved U12 splicing mechanisms between maize and humans with both RBM48 and ARMC7 having roles in the activated spliceosome.

maize↗

AtDGCR14L contributes to salt-stress tolerance via regulating pre-mRNA splicing in Arabidopsis

In plants, the pre-mRNA alternative splicing has been demonstrated to be a crucial tier that regulates gene expression in response to salt stress. However, the underlying mechanisms remain elusive. Here, in this study, we studied the roles of DIGEORGE-SYNDROME CRITICAL REGION 14-like (AtDGCR14L) in regulating pre-mRNA splicing and salt stress tolerance. We discovered that Arabidopsis AtDGCR14L is required for maintaining plant salt stress tolerance and the constitutively spliced and active isoforms of important stress- and/or abscisic acid (ABA)-responsive genes. We also identified the interaction between AtDGCR14L and splicing factor U1-70k, which needs a highly conserved three amino acid (TWG) motif in DGCR14. Different from wild-type AtDGCR14L, the overexpression of TWG-substituted AtDGCR14L mutant did not change salt stress tolerance or pre-mRNA splicing of stress/ABA-responsive genes. Additionally, SWITCH3A (SWI3A) is a core subunit of the SWI/SUCROSE NONFERMENTING (SWI/SNF) chromatin-remodeling complexes. We found that SWI3A, whose splicing depends on AtDGCR14L, actively enhances salt stress tolerance. These results revealed that AtDGCR14L may play an essential role in crosstalk between plant salt-stress response and pre-mRNA splicing mechanisms. We also unveiled the potential role of SWI3A in controlling salt stress tolerance. The TWG motif in the intrinsically disordered region of AtDGCR14L is highly conserved and crucial for DGCR14 functions.

59 BASIC BIOLOGICAL SCIENCES↗

An interpretable model of pre-mRNA splicing for animal and plant genes

Pre-mRNA splicing is a fundamental step in gene expression, conserved across eukaryotes, in which the spliceosome recognizes motifs at the 3' and 5' splice sites (SSs), excises introns, and ligates exons. SS recognition and pairing is often influenced by protein splicing factors (SFs) that bind to splicing regulatory elements (SREs). Here, we describe SMsplice, a fully interpretable model of pre-mRNA splicing that combines models of core SS motifs, SREs, and exonic and intronic length preferences. We learn models that predict SS locations with 83 to 86% accuracy in fish, insects, and plants and about 70% in mammals. Learned SRE motifs include both known SF binding motifs and unfamiliar motifs, and both motif classes are supported by genetic analyses. Our comparisons across species highlight similarities between non-mammals, increased reliance on intronic SREs in plant splicing, and a greater reliance on SREs in mammalian splicing.

59 BASIC BIOLOGICAL SCIENCES↗

Selected humanization of yeast U1 snRNP leads to global suppression of pre-mRNA splicing and mitochondrial dysfunction in the budding yeast

The recognition of the 5′ splice site (5′ ss) is one of the earliest steps of pre-mRNA splicing. To better understand, the mechanism and regulation of 5′ ss recognition, we selectively humanized components of the yeast U1 (yU1) snRNP to reveal the function of these components in 5′ ss recognition and splicing. We targeted U1C and Luc7, two proteins that interact with and stabilize the yU1 snRNA and the 5′ ss RNA duplex. We replaced the zinc-finger (ZnF) domain of yeast U1C (yU1C) with its human counterpart, which resulted in a cold-sensitive growth phenotype and moderate splicing defects. We next added an auxin-inducible degron to yeast Luc7 (yLuc7) protein (to mimic the lack of Luc7Ls in human U1 snRNP). We found that Luc7-depleted yU1 snRNP resulted in the concomitant loss of Prp40 and Snu71 (two other essential yU1 snRNP proteins), and further biochemical analyses suggest a model of how these three proteins interact with each other in the U1 snRNP. The loss of these proteins resulted in a significant growth retardation accompanied by a global suppression of pre-mRNA splicing. The splicing suppression led to mitochondrial dysfunction as revealed by a release of Fe 2+ into the growth medium and an induction of mitochondrial reactive oxygen species. Together, these observations indicate that the human U1C ZnF can substitute that of yeast, Luc7 is essential for the incorporation of the Luc7–Prp40–Snu71 trimer into yU1 snRNP, and splicing plays a major role in the regulation of mitochondrial function in yeast.

Biochemistry & Molecular Biology↗

Thermally robust and highly stable method for splicing silica glass fiber to crystalline sapphire fiber

This research reports an advancement in splicing silica glass fiber to sapphire single-crystal optical fiber (SCF) using a specialized glass processing device, including data that demonstrate the thermal stability of the splice to 1000°C. A filament heating process was used to produce a robust splice between the dissimilar fibers. A femtosecond laser is used to inscribe a fiber Bragg gratings sensor into the SCF to measure the high-temperature capabilities and signal attenuation characteristics of the splice joint. The experimental results demonstrate that the proposed splicing method produces a splice joint that is robust, stable, repeatable, and withstands temperatures up to 1000°C with a low attenuation of 0.5 dB. Here, the proposed method allows placement of SCF-based sensors in the extreme environments encountered in various engineering fields, such as nuclear, chemical, aviation, and metals manufacturing, to enable improvements in process monitoring, product quality, and production efficiency.

36 MATERIALS SCIENCE↗

The behavior of bonded doubler splices for composite sandwich panels

The results of an investigation into the behavior of adhesively bonded doubler splices of two composite material sandwich panels are presented. The splices are studied from three approaches: analytical; numerical (finite elements); and experimental. Several parameters that characterize the splice are developed to determine their influence upon joint strength. These parameters are: doubler overlap length; core stiffness; laminate bending stiffness; the size of the gap between the spliced sandwich panels; and room and elevated temperatures. Similarities and contrasts between these splices and the physically similar single and double lap joints are discussed. The results of this investigation suggest several possible approaches to improving the strength of the sandwich splices.

Zeller, T. A.↗

NASA SPLICE Project: Development and Testing of Precision Landing GN&C Technologies

NASA's technology advancement needs for entry, descent and landing call for high-precision, high-rate sensors that can improve navigation accuracy and vehicle control performance. Higher landing accuracy is required for any future human lander missions, and likely, for most robotic missions 1,2. Sensors and algorithms that significantly reduce navigation errors and can image the local terrain will enable landing at locations of high scientific interest that would otherwise pose significant risk to the vehicle. The Safe and Precise Landing-Integrated Capabilities Evolution project, or SPLICE, is developing precision landing and hazard avoidance (PL&HA) technologies for NASA and for potential commercial space flight missions. SPLICE technologies include sensors, algorithms, advanced space flight computing capabilities, and simulation tools used to integrate and study guidance, navigation, and control (GN&C) system performance. SPLICE efforts include hardware-in-the-loop (HWIL) simulation testing, ground testing, and flight testing, including reuse of hardware from the CoOperative Blending of Autonomous Landing Technologies (COBALT) suborbital flight-test payload3,4. Two of the precise navigation sensors that are being developed and matured within SPLICE are LiDARs. Since 2006, NASA Langley has been developing a Navigation Doppler LiDAR (NDL) for precise velocity measurements, and SPLICE is building an NDL engineering test unit (ETU) that will be brought up to TRL 6 following environmental and high-speed1,2 testing. NASA Goddard is developing a Hazard Detection LiDAR (HD LiDAR) engineering development unit (EDU) for SPLICE that has relevance to future human and robotic lander missions. The HD LiDAR will be flight test and matured to TRL 5.

Restrepo, Carolina I.↗

A System to Provide Deterministic Flight Software Operation and Maximize Multicore Processing Performance: The Safe and Precise Landing – Integrated Capabilities Evolution (SPLICE) Datapath

A method and design are described for a system that processes multiple data streams, utilizing a multicore asymmetric processing architecture, that eliminates data interrupts to the application processors. The design supports a deterministic environment for flight software in NASA’s Safe and Precise Landing – Integrated Capabilities Evolution (SPLICE) project. The SPLICE project develops sensor, algorithm, and compute technologies for Precision Landing and Hazard Avoidance (PL&HA) capabilities. The compute technology for SPLICE is the Descent and Landing Computer (DLC). The DLC hosts several SPLICE algorithms with high computational resource requirements that must be executed in a real-time and deterministic manner. The software runs on a custom Single Board Computer (SBC), with a Xilinx Ultrascale+ Multiprocessor System-on-a-Chip (MPSoC). Input data for the flight software is from a variety of sensors, unique with respect to data rate and packet size. A data path between the SPLICE sensors and algorithms is designed to efficiently deliver this data to the flight software using the MPSoC asymmetric processing cores and Field Programmable Gate Array (FPGA) fabric. This is implemented in a manner that isolates the application processors running the flight software from interrupts associated with the input data. By leveraging real-time processors on the MPSoC, and a structure with the appropriate interfaces in the shared memory on the SBC, the flight software can use the full set of application processors. The available utilization for each processor in this set is also maximized for the SPLICE applications, providing a sufficiently deterministic execution environment without the cost and overhead of a real-time operating system.

heterogeneous processing system↗

A System to Provide Deterministic Flight Software Operation and Maximize Multicore Processing Performance: The Safe and Precise Landing – Integrated Capabilities Evolution (SPLICE) Datapath

A method and design are described for a system that processes multiple data streams, utilizing a multicore asymmetric processing architecture, that eliminates data interrupts to the application processors. The design supports a deterministic environment for flight software in NASA’s Safe and Precise Landing – Integrated Capabilities Evolution (SPLICE) project. The SPLICE project develops sensor, algorithm, and compute technologies for Precision Landing and Hazard Avoidance (PL&HA) capabilities. The compute technology for SPLICE is the Descent and Landing Computer (DLC). The DLC hosts several SPLICE algorithms with high computational resource requirements that must be executed in a real-time and deterministic manner. The software runs on a custom Single Board Computer (SBC), with a Xilinx Ultrascale+ Multiprocessor System-on-a-Chip (MPSoC). Input data for the flight software is from a variety of sensors, unique with respect to data rate and packet size. A data path between the SPLICE sensors and algorithms is designed to efficiently deliver this data to the flight software using the MPSoC asymmetric processing cores and Field Programmable Gate Array (FPGA) fabric. This is implemented in a manner that isolates the application processors running the flight software from interrupts associated with the input data. By leveraging real-time processors on the MPSoC, and a structure with the appropriate interfaces in the shared memory on the SBC, the flight software can use the full set of application processors. The available utilization for each processor in this set is also maximized for the SPLICE applications, providing a sufficiently deterministic execution environment without the cost and overhead of a real-time operating system.

David K. Rutishauser↗

Sample Processor for Life on Icy Worlds (SPLIce): Design and Test Results

We report the design, development, and testing of the Sample Processor for Life on Icy Worlds (SPLIce) system, a microfluidic sample processor to enable autonomous detection of signatures of life and measurements of habitability parameters in Ocean Worlds. This monolithic fluid processing-and-handling system (Figure 1; mass 0.5 kg) retrieves a 50-L-volume sample and prepares it to supply a suite of detection instruments, each with unique preparation needs. SPLIce has potential applications in orbiter missions that sample ocean plumes, such as found in Saturns icy moon Enceladus, or landed missions on the surface of icy satellites, such as Jupiters moon Europa. Answering the question Are we alone in the universe? is captivating and exceptionally challenging. Even general criteria that define life very broadly include a significant role for water [1,2]. Searches for extinct or extant life therefore prioritize locations of abundant water whether in ancient (Mars), or present (Europa and Enceladus) times. Only two previous planetary missions had onboard fluid processing: the Viking Biology Experiments [3] and Phoenixs Wet Chemistry Laboratory (WCL) [4]. SPLIce differs crucially from those systems, including its capability to process and distribute L-volume samples and the integration autonomous control of a wide range of fluidic functions, including: 1) retrieval of fluid samples from an evacuated sample chamber; 2) onboard multi-year storage of dehydrated reagents; 3) integrated pressure, pH, and conductivity measurement; 4) filtration and retention of insoluble particles for microscopy; 5) dilution or vacuum-driven concentration of samples to accommodate instrument working ranges; 6) removal of gas bubbles from sample aliquots; 7) unidirectional flow (check valves); 8) active flow-path selection (solenoid-actuated valves); 9) metered pumping in 100 nL volume increments. The SPLIce manifold, made of three thermally fused layers of precision-machined cyclo-olefin polymer, supports all fluidic components (Figure 1) and integrated microchannels (125 x 250 m). Fluid is pumped by a stepper-motor-driven pump (Lee Co.). The functionality of the integrated MEMS pressure sensor (Honeywell) and passive check valves (Figure 2) were tested in conjunction with our newly designed integral bubble traps (Figure 3) and hydrophobic membrane-based concentrator (Figure 4). The concentrator (initially tested as a standalone component) demonstrated 5-fold vacuum-evaporative concentration. Polyethylene fused bead beds (PEFBBs; 50 porosity) store drylyophilized buffers, calibrants, and fluorescent dyes, and also promote mixing of sample with calibrant, dye, or H2O. Software-controlled automated tests demonstrated successful 1) fluid delivery to each component 2) valve and pump synchronization 3) sample aliquot delivery to instrument interface ports, and 4) rehydration of vacuum-dried fluorescent dye. In Figure 5, fluorescein on PEFBBs was rehydrated for 15 min using a pump-delivered water aliquot; it is displaced as H2O enters the bottom of the channel and pushes the dye into a check valve. Ultimately, SPLIce will fluorescently label amino acids in the sample for microchip-based electrophoretic (MCE) chiral separation and detection to seek and quantify key organic bio-signatures [5]; it will also deliver sample to a microfluidic version of WCL (mWCL) to measure soluble ions and redox-active species.

Life detection↗

Visualizing group II intron dynamics between the first and second steps of splicing

Group II introns are ubiquitous self-splicing ribozymes and retrotransposable elements evolutionarily and chemically related to the eukaryotic spliceosome, with potential applications as gene-editing tools. Recent biochemical and structural data have captured the intron in multiple conformations at different stages of catalysis. Here, we employ enzymatic assays, X-ray crystallography, and molecular simulations to resolve the spatiotemporal location and function of conformational changes occurring between the first and the second step of splicing. We show that the first residue of the highly-conserved catalytic triad is protonated upon 5’-splice-site scission, promoting a reversible structural rearrangement of the active site (toggling). Protonation and active site dynamics induced by the first step of splicing facilitate the progression to the second step. Our insights into the mechanism of group II intron splicing parallels functional data on the spliceosome, thus reinforcing the notion that these evolutionarily-related molecular machines share the same enzymatic strategy.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

The Characteristics of Fatigue Damage in the Fuselage Riveted Lap Splice Joint

An extensive data base has been developed to form the physical basis for new analytical methodology to predict the onset of widespread fatigue damage in the fuselage lap splice joint. The results of detailed destructive examinations have been cataloged to describe the physical nature of MSD in the lap splice joint. ne catalog includes a detailed description, e.g., crack initiation, growth rates, size, location, and fracture morphology, of fatigue damage in the fuselage lap splice joint structure. Detailed examinations were conducted on a lap splice joint panel removed from a full scale fuselage test article after completing a 60,000 cycle pressure test. The panel contained a four bay region that exhibited visible outer skin cracks and regions of crack link-up along the upper rivet row. Destructive examinations revealed undetected fatigue damage in the outer skin, inner skin, and tear strap regions. Outer skin fatigue cracks were found to initiate by fretting damage along the faying surface. The cracks grew along the faying surface to a length equivalent to two to three skin thicknesses before penetrating the outboard surface of the outer skin. Analysis of fracture surface marker bands produced during full scale testing revealed that all upper rivet row fatigue cracks contained in a dim bay region grow at similar rates; this important result suggests that fracture mechanics based methods can be used to predict the growth of outer skin fatigue cracks in lap splice structure. Results are presented showing the affects of MSD and out-of-plane pressure loads on outer skin crack link-up.

Piascik, Robert S.↗