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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 325 records · Page 18
All sourcesNASA NTRSDOE OSTI
NASA NTRS2005

SAFER Inspection of Space Shuttle Thermal Protection System

In the aftermath of the space shuttle Columbia accident, it quickly became clear that new methods would need to be developed that would provide the capability to inspect and repair the shuttle's thermal protection system (TPS). A boom extension to the Remote Manipulator System (RMS) with a laser topography sensor package was identified as the primary means for measuring the damage depth in acreage tile as well as scanning Reinforced Carbon- Carbon (RCC) surfaces. However, concern over the system's fault tolerance made it prudent to investigate alternate means of acquiring close range photographs and contour depth measurements in the event of a failure. One method that was identified early was to use the Simplified Aid For EVA Rescue (SAFER) propulsion system to allow EVA access to damaged areas of concern. Several issues were identified as potential hazards to SAFER use for this operation. First, the ability of an astronaut to maintain controlled flight depends upon efficient technique and hardware reliability. If either of these is insufficient during flight operations, a safety tether must be used to rescue the crewmember. This operation can jeopardize the integrity of the Extra-vehicular Mobility Unit (EMU) or delicate TPS materials. Controls were developed to prevent the likelihood of requiring a tether rescue, and procedures were written to maximize the chances for success if it cannot be avoided. Crewmember ability to manage tether cable tension during nominal flight also had to be evaluated to ensure it would not negatively affect propellant consumption. Second, although propellant consumption, flight control, orbital dynamics, and flight complexity can all be accurately evaluated in Virtual Reality (VR) Laboratory at Johnson Space Center, there are some shortcomings. As a crewmember's hand is extended to simulate measurement of tile damage, it will pass through the vehicle without resistance. In reality, this force will push the crewmember away from the vehicle, and could induce a moment which, if strong enough, could saturate the attitude control system in SAFER. This raises the concern that additional propellant will be consumed to maintain controlled flight. To account for this, the fidelity of the Virtual Reality simulation was improved to include the effect of crewmember contact with the vehicle during SAFER flight. In addition, while participating in VR simulations, the subject is in shirt sleeves and sits in a chair. This does not provide a flight-like representation of body position awareness. To prevent inadvertent contact with tile or RCC, other facilities were utilized to establish crew preferences for body attitude and tool configuration. Finally, a study was performed to determine if attitude constraints are needed for the Space shuttle and International Space Station to reduce SAFER flight difficulty.

Scoville, Zebulon C.↗
NASA NTRS2008

Sea State and Weather Capability for NASA's Constellation Program

Marine weather and related parameters such as wind, ocean wave height and period, air temperature, sea surface temperature, visibility, and potential for icing are critical to the design, operation, and safety of crewed space vehicles. The National Aeronautics and Space Administration's (NASA's) Constellation Program requires detailed assessment of marine weather related parameters that may be encountered during launch, abort, landing, and crew rescue operations for the crewed Axes/Orion space vehicles. This information is required for both space vehicle design and operational purposes. The space vehicles must be designed such that they cam withstand the environment they are likely to encounter. The crewed Axes/Orion space vehicles will launch from NASA's Kennedy Space Center (KSC), Florida for both International Space Station (ISS) missions with 51.6deg inclination orbits and lunar missions with approximately 280 inclination orbits. Since both missions will fly ever the Atlantic Ocean on ascent to orbit and will fly over the Pacific Ocean on descent from orbit, an unlikely but possible emergency abort could require parachuting the Orion capsule and crew into the ocean. This situation could potentially put the crew in an isolated and hazardous environment for severn hours while they await rescue. Therefore, abort, landing, and crew rescue elements of the Constellation Program must address weather related parameters on a global scale. This paper describes buoy measurement data, sea surface temperature satellite data, and sea state computer model data that are being utilized by the Constellation Program to address these design and operational issues.

Barbre, R. E.↗
NASA NTRS2008

Sea State and Weather Assessment Capability for NASA's Constellation Program

Marine weather and related parameters such as wind, ocean wave height and period, air temperature, sea surface temperature, visibility, and potential for icing are critical to the design, operation, and safety of crewed space vehicles. The National Aeronautics and Space Administration's (NASA's) Constellation Program requires detailed assessment of marine weather related parameters that may be encountered during launch, abort, landing, and crew rescue operations for the crewed Ares/Orion space vehicles. This information is required for both space vehicle design and operational purposes. The space vehicles must be designed such that they can withstand the environment they are likely to encounter. The crewed Ares/Orion space vehicles will launch from NASA's Kennedy Space Center (KSC), Florida for both International Space Station (ISS) missions with 51.6 o inclination orbits and lunar missions with approximately 28 inclination orbits. Since both missions will fly over the Atlantic Ocean on ascent to orbit and will fly over the Pacific Ocean on descent from orbit, an unlikely but possible emergency abort could require parachuting the Orion capsule and crew into the ocean. This situation could potentially put the crew in an isolated and hazardous environment for several hours while they await rescue. Therefore, abort, landing, and crew rescue elements of the Constellation Program must address weather related parameters on a global scale. This paper describes buoy measurement data, sea surface temperature satellite data, and sea state computer model data that are being utilized by the Constellation Program to address these design and operational issues.

Barbre, B. J.↗
NASA NTRS2003

Modeled Microgravity-Induced Protein Kinase C Isoform Expression in Human Lymphocytes

In long-term space travel, the crew is exposed to microgravity and radiation that invoke potential hazards to the immune system. T cell activation is a critical step in the immune response. Receptor-mediated signaling is inhibited both in microgravity and modeled microgravity (MMG) as reflected in diminished DNA synthess in peripheral blood lymphocytes and their locomotion through gelled type 1 collagen. Direct activation of Protein Kinase C (PKC) bypassing cell surface events using the phorbol ester PMA rescues MMG-inhibited lymphocyte activation and locomotion, whereas calcium ionophore ionomycin had no rescue effect. Thus calcium-independent PKC isoforms may be affected in MMG-induced locomotion inhibition and rescue. Both calcium-dependent isoforms and calcium-independent PKC isoforms were investigated to assess their expression in lymphocytes in 19 and MMG-culture. Human lymphocytes were cultured and harvested at 24, 48, 72 and 96 hours and serial samples assessed for locomotion using type I collagen and expression of PKC isoforms. Expression of PKC-alpha, -delta and -epsilon was assessed by RT-PCR, flow cytometry and immunoblotting. Results indicated that PKC isoforms delta and epsilon were down-regulated by more than 50% at the transcriptional and translational levels in MMG-cultured lymphocytes compared with 19 controls. Events upstream of PKC such as phosphorylation of Phospholipase C(gamma) (PLC-gamma) in MMG, revealed accumulation of inactive enzyme. Depressed Ca++ -independent PKC isoforms may be a consequence of an upstream lesion in the signal transduction pathway. The differential response among calcium-dependent and calcium-independent isoforms may actually result from MMG intrusion events earlier than, but after ligand-receptor interaction. Keywords: Signal transduction, locomotion, immunity

Sundaresan, A.↗
NASA NTRS2012

13 Things That Saved Apollo 13

Perhaps, the most exciting rescue, terrestrial or extra-terrestrial, is the successful return of the Apollo 13 crew to Earth in April of 1970. The mission s warning system engineer, Jerry Woodfill, who remains a NASA employee after 47 years of government service has examined facets of the rescue for the past 42 years. He will present "13 Things That Saved Apollo 13" from the perspective of his real time experience as well as two score years of study. Many are recent discoveries never before published in mission reports, popular books or documentary and Hollywood movies depicting the rescue.

Woodfill, Jared↗
NASA NTRS2012

Hands-Free Control Interfaces for an Extra Vehicular Jetpack

The National Aeronautics and Space Administration (NASA) strategic vision includes, as part of its long-term goals, the exploration of deep space and Near Earth Asteroids (NEA). To support these endeavors, funds have been invested in research to develop advanced exploration capabilities. To enable the human mobility necessary to effectively explore NEA and deep space, a new extravehicular activity (EVA) Jetpack is under development at the Johnson Space Center. The new design leverages knowledge and experience gained from the current astronaut rescue device, the Simplified Aid for EVA Rescue (SAFER). Whereas the primary goal for a rescue device is to return the crew to a safe haven, in-space exploration and navigation requires an expanded set of capabilities. To accommodate the range of tasks astronauts may be expected to perform while utilizing the Jetpack, it was desired to offer a hands-free method of control. This paper describes the development and innovations involved in creating two hands-free control interfaces and an experimental test platform for a suited astronaut flying the Jetpack during an EVA.

Zumbado, Jennifer Rochlis↗
NASA NTRS2016

Preserving, Enhancing, and Continuing the Scientific Legacy of the Apollo Sample Suite

From 1969 to 1972, Apollo astronauts collected 382 kg of rocks, soils, and core samples from six geologically diverse locations on the Moon. In the nearly 50 years since the samples were collected, over 3000 different studies have been conducted using the nearly 2200 different Apollo samples. Despite the maturity of the sample collection, many new studies of lunar samples are undertaken each year, with an average of more than 55 requests and more than 600 distinct subsamples allocated annually over the past five years. The Apollo samples are a finite resource, however. Although new studies are encouraged, it is important that new studies do not duplicate previous studies, and where possible, leverage previous results to inform and enhance the current studies. This helps to preserve the samples and scientific funding, both of which are precious resources. We have initiated several new efforts to rescue some of the early analyses from these samples, including unpublished analytical data. We are actively scanning NASA documentation in paper form that is related to the Apollo missions and sample processing, and we are collaborating with IEDA to establish a geochemical data base called MoonDB. To populate this database, we are actively working with about a dozen prominent lunar PIs to organize and transcribe years of both published and unpublished data, making it available to all researchers. This effort will also take advantage of new online analytical tools like PetDB. There have already been tangible results from the MoonDB data rescue effort. A pilot project involving the rescue of geochemical data of John Delano on Apollo pyroclastic glasses has already been referenced in multiple Apollo sample requests, and in fact, the compiled data was used as part of one of the new studies. Similarly, scanned sample handling reports have been utilized to find previously analyzed samples that were appropriate to fulfill new sample requests. We have also begun to image the Apollo samples using (1) micro‐CT scanning to document the interior structure of samples, and (2) comprehensive high resolution photography of samples, enabling high resolution 3D reconstructions of the samples. Both efforts will provide comprehensive access to these samples and allow for more targeted requests, and thus better curation of the samples for decades to come.

Zeigler, Ryan↗
NASA NTRSDate not supplied

The Wizard Staff: A Mobile Tool Supporting Human Exploration on The Lunar Surface

The Wizard Staff is a rechargeable, battery powered mobile tool with multiple uses for lunar surface exploration. It supports human activity as a surface survey tool, provides external illumination to enhance safety, delivers networking, communications, and navigation features to remote sites, and assists crew rescue with an emergency beacon and rigid splint structure. As a survey tool, it assists crewed geologic field site exploration with external lights, cameras, and LiDAR to illuminate, map, and record the sample collection process. As a relay station, it provides a surface intranet communications and navigation node for extended exploration far from the lander or habitation module. A separately developed Search-and-Rescue (SAR) handheld device can broadcast a signal to the Gandalf Staff containing crew position and biometric data. The staff then formats the message and sends it orbiting satellite(s) for emergency response via the LunaNET on a dedicated frequency similar to terrestrial applications using an Emergency Position-Indicating Radiobeacon (EPIRB). The staff is designed with an upper and lower segment. Components in the upper segment are the LiDAR, external lighting arrays, cameras, communications equipment, intelligence devices, and user interface for crew operations. The lower segment is a set of FeLiPO4 battery cells packed into the 2” diameter carbon fiber tube. When the battery pack is discharged, a spring connector (designed for the dusty lunar environment) easily disconnects the two segments for replacement with another fully charged battery pack. The lower segment can be used for Incapacitated Crew Rescue (ICR) as a rigid splint. As a stand-alone science platform, the Wizard Staff provides infrastructure for instruments and sensors to record the lunar environment over long periods of time. After a crew installs the components on the lunar surface, the platform delivers data to the lunar lander or base station for analysis. The stand-alone mode requires an auxiliary power source (e.g. solar array) and energy storage system (e.g. battery), so it could also be used as an electrical recharging station. A working prototype of Wizard Staff has been developed with collaboration between NASA (JSC, MSFC, and GSFC), Texas Space Technology Applications and Research (T STAR), and Texas A&M University (TAMU) using $300k of innovation funding in FY’21 and FY’22. The functional staff weights 36# on Earth (6# on the Moon), stands 2.1m tall and delivers external lighting with an array of 40 LED bulbs generating 1460 Lux at 2m range (near field), or 2 Lux at 50m (far field).

Gandalf Staff↗
NASA NTRSDate not supplied

EVA Planning: Using Neutral Buoyancy Laboratory (NBL) Training to Predict in-Flight Energy Expenditure

Metabolic rate (“met rate”) is the amount of energy expended over a period of time and is influenced by many factors including body composition, level of physical activity, resting metabolic rate, sex, age, and food intake. Met rate is measured during Extravehicular Activity (EVA) training at the Neutral Buoyancy Laboratory (NBL) and during in-flight EVAs through indirect calorimetry, calculating energy expenditure from respiratory measurements of O 2 consumption and/or CO 2 production. During Extravehicular Activity (EVA) planning, metabolic cost is important to consider and is used to inform EVA duration based on spacesuit consumables associated with life support systems. Currently, NBL and previous ISS EVA met rate data for specified crewmembers are utilized to predict in-flight EVA metabolic costs based on a proposed EVA timeline. Timeline data collected during training is used to relate met rates to specific EVA activities, which are in turn assigned to more generalized EVA task categories, categorizing by both task type and restraint type. EVA task categories include EVA Setup/Cleanup, Worksite Setup/Cleanup, Cable Routing, Bolts, Fluid Connectors, Electrical Connectors, R&R Work, Miscellaneous Work, Incapacitated Crew Rescue (being rescued or performing), Assisted Crew Rescue (being assisted or performing), and Translation. Restraint types consist of Free-Float, Body Restraint Tether (BRT), Articulating Portable Foot Restraint (APFR), and Space Station Remote Manipulator System (SSRMS). From a crewmember’s historical data, individualized 10th, 50th and 90th percentile met rate estimates are generated for each task category and used to estimate the proposed EVA timeline metabolic cost. In-flight metabolic data (“As-Executed”) from recent ISS US EVAs 85-88 (totaling eight EVA crewmember met rates) was compared with their predicted metabolic cost (“As-Planned”) to evaluate the accuracy of the current met rate estimation method. Across the four EVAs, As-Executed Cumulative EVA Total Metabolic Cost (M = 5893.78 BTU, SD = 816.60) was not significantly different compared to As-Planned Cumulative EVA Total Metabolic Cost (M = 6106.60 BTU, SD = 826.62; t(7) = 0.751 , p = .477). Though not a significant difference, generally, As-Planned total estimates were slightly higher than As-Executed total metabolic cost. Relative Error for Cumulative EVA Total Metabolic Cost ranged from -33% to 14.7%, depending on the crewmember and EVA. When comparing As-Planned to A-Executed EVA task categories for Bolts, Electrical Connectors, EVA Cleanup, EVA Setup, Miscellaneous Work, Translation, Worksite Cleanup, and Worksite Setup during these EVAs, no significant differences were observed, however, there was a significant difference in As-Planned (M = 783.71 BTU, SD = 408.88) compared to As-Executed (M = 608.87 BTU, SD = 425.29) metabolic cost for the task category of Repair-and-Replace (R&R) Work (t(17) = 3.21 , p = .005). Looking closer within the R&R Work task category, As-Executed R&R Work with Free-Float restraint type (M = 706.17 BTU, SD = 352.18) was significantly less than As-Planned R&R Work with Free-Float restraint type (M= 887.96 BTU, SD = 381.42; t(12) = 2.59, p < .024). As-Executed R&R Work with SSRMS Restraint type (M = 355.89 BTU, SD = 534.65) was not significantly different from As-Planned values (M = 512.69 BTU, SD = 383.34; t(4) = 1.88, p = 0.132). These findings suggest that the energy expended performing R&R Work (Free-Float) is lower in flight than predicted. Accurate predictions of the metabolic cost of EVA are essential for planning and executing successful ISS EVAs. Overall, the current met rate prediction method is similar to actual in-flight values, slightly erring on the side of overestimation. Future work includes analysis of more historical in-flight EVA data to increase the power of the analysis, evaluating the NBL-ISS met rate conversion factor between NBL and ISS tasks, as well as exploring methods of substitution when crewmembers are missing prior task category data.

Lauren Cox↗
DOE OSTI2014

Increasing the structural coverage of tuberculosis drug targets

High-resolution three-dimensional structures of essential Mycobacterium tuberculosis (Mtb) proteins provide templates for TB drug design, but are available for only a small fraction of the Mtb proteome. Here we evaluate an intra-genus “homolog-rescue” strategy to increase the structural information available for TB drug discovery by using mycobacterial homologs with conserved active sites. We found that of 179 potential TB drug targets selected for x-ray structure determination, only 16 yielded a crystal structure. By adding 1675 homologs from nine other mycobacterial species to the pipeline, structures representing an additional 52 otherwise intractable targets were solved. To determine whether these homolog structures would be useful surrogates in TB drug design, we compared the active sites of 106 pairs of Mtb and non-TB mycobacterial (NTM) enzyme homologs with experimentally determined structures, using three metrics of active site similarity, including superposition of continuous pharmacophoric property distributions. Pair-wise structural comparisons revealed that 19/22 pairs with >55% overall sequence identity had active site Cα RMSD <1 Å, >85% side chain identity, and ≥80% PS APF (similarity based on pharmacophoric properties) indicating highly conserved active site shape and chemistry. Applying these results to the 52 NTM structures described above, 41 shared >55% sequence identity with the Mtb target, thus increasing the effective structural coverage of the 179 Mtb targets over three-fold (from 9% to 32%). The utility of these structures in TB drug design can be tested by designing inhibitors using the homolog structure and assaying the cognate Mtb enzyme; a promising test case, Mtb cytidylate kinase, is described. The homolog-rescue strategy evaluated here for TB is also generalizable to drug targets for other diseases.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

DNA Sequence Analysis of an Inversion Hot Spot in Lobeliaceae Plastomes

The evolution of plastid genomes (plastomes) in land plants is typically conservative, with extensive structural rearrangements present in only a few groups. Early Southern blot analysis identified two Lobelia species that minimally required deletion of the plastid gene accD and five inversions to account for their plastome arrangement relative to the ancestral organization. Sixty alternative 5-step inversion scenarios could account for the observed arrangement, but only one scenario was consistent with the criterion of ‘common cause’ attributable to a putative rearrangement hot spot at the accD deletion-site. Plastome sequencing demonstrated that this previously hypothesized inversion order is historically accurate. Detailed reconstructions of the ancestral plastome organization before and after each inversion are presented herein. Stem-loop and disruption-rescue models were evaluated for each inversion. One inversion has an obvious stem-loop basis, but the other four inversions were primarily caused by serial insertion of foreign (extra-plastid) DNA bearing large open-reading frames that disrupted plastome organization at the accD deletion-site, and complete plastomes were rescued by seemingly arbitrary ligation or fortuitous recombination at the other inversion endpoint. Transposed copies of DNA segments from elsewhere in the plastome are frequently inserted at inversion junctions, and four junctions are consistent with the stem-loop ligation model.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Expression and purification of affinity-tagged variants of the photochemical reaction center from Heliobacterium modesticaldum

The heliobacterial photochemical reaction center (HbRC) from the chlorophototrophic Firmicutes bacterium Heliobacterium modesticaldum is the only homodimeric type I RC whose structure is known. Using genetic techniques recently established in our lab, we have developed a rapid heterologous expression system for the HbRC core polypeptide PshA. Our system relies on rescue of the non-chlorophototrophic ΔpshA::cbp2p - aph3 strain of Hbt. modesticaldum by expression of a heterologous pshA gene from a replicating shuttle vector. Furthermore, we constructed two tagged variants of PshA, one with an N-terminal octahistidine tag and one with an internal hexahistidine tag, which facilitate rapid purification of pure, active HbRC cores in milligram quantities. We constructed a suite of shuttle vectors bearing untagged or tagged versions of pshA driven by various promoters. Surprisingly, we found that the eno and gapDH_2 promoters from Clostridium thermocellum drive better expression of pshA than fragments of DNA derived from the region upstream of the pshA locus on the Hbt. modesticaldum genome. This “ pshA rescue” strategy additionally provided a useful window into how Hbt. modesticaldum regulates pigment synthesis and growth rate when chlorophototrophic output decreases.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

The mitochondrially targeted peptide elamipretide (SS-31) improves ADP sensitivity in aged mitochondria by increasing uptake through the adenine nucleotide translocator (ANT)

Aging muscle experiences functional decline in part mediated by impaired mitochondrial ADP sensitivity. Elamipretide (ELAM) rapidly improves physiological and mitochondrial function in aging and binds directly to the mitochondrial ADP transporter ANT. We hypothesized that ELAM improves ADP sensitivity in aging leading to rescued physiological function. We measured the response to ADP stimulation in young and old muscle mitochondria with ELAM treatment, in vivo heart and muscle function, and compared protein abundance, phosphorylation, and S-glutathionylation of ADP/ATP pathway proteins. ELAM treatment increased ADP sensitivity in old muscle mitochondria by increasing uptake of ADP through the ANT and rescued muscle force and heart systolic function. Protein abundance in the ADP/ATP transport and synthesis pathway was unchanged, but ELAM treatment decreased protein s-glutathionylation incuding of ANT. Mitochondrial ADP sensitivity is rapidly modifiable. This research supports the hypothesis that ELAM improves ANT function in aging and links mitochondrial ADP sensitivity to physiological function.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2010

Over-expression of COQ10 in Saccharomyces cerevisiae inhibits mitochondrial respiration

Research highlights: {yields} COQ10 deletion elicits a defect in mitochondrial respiration correctable by addition of coenzyme Q{sub 2}, a synthetic diffusible ubiquinone. {yields} The significance that purified Coq10p contains bound Q{sub 6} was examined by testing over-expression of Coq10p on respiration. {yields} Inhibition of CoQ function due to Coq10p excess strength our hypothesis of Coq10p function in CoQ delivery. {yields} Respiratory deficiency caused by more Coq10p was specific and restored by Q{sub 2} in mitochondria or by Coq8p in cells. {yields} Coq8p over-production on other coq mutants revealed a surprisingly higher stability of other Coq proteins. -- Abstract: COQ10 deletion in Saccharomyces cerevisiae elicits a defect in mitochondrial respiration correctable by addition of coenzyme Q{sub 2}. Rescue of respiration by Q{sub 2} is a characteristic of mutants blocked in coenzyme Q{sub 6} synthesis. Unlike Q{sub 6} deficient mutants, mitochondria of the coq10 null mutant have wild-type concentrations of Q{sub 6}. The physiological significance of earlier observations that purified Coq10p contains bound Q{sub 6} was examined in the present study by testing the in vivo effect of over-expression of Coq10p on respiration. Mitochondria with elevated levels of Coq10p display reduced respiration in the bc1 span of the electron transport chain, which can be restored with exogenous Q{sub 2}. This suggests that in vivo binding of Q{sub 6} by excess Coq10p reduces the pool of this redox carrier available for its normal function in providing electrons to the bc1 complex. This is confirmed by observing that extra Coq8p relieves the inhibitory effect of excess Coq10p. Coq8p is a putative kinase, and a high-copy suppressor of the coq10 null mutant. As shown here, when over-produced in coq mutants, Coq8p counteracts turnover of Coq3p and Coq4p subunits of the Q-biosynthetic complex. This can account for the observed rescue by COQ8 of the respiratory defect in strains over-producing Coq10p.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2010

Corepressor MMTR/DMAP1 is an intrinsic negative regulator of CAK kinase to regulate cell cycle progression

Research highlights: {yields} Co-repressor MMTR/DMAP1 is an intrinsic negative regulator of CAK kinase. {yields} MMTR inhibited cell proliferation due to delays of G1/S and G2/M transitions. {yields} Co-expression of MAT1 and MMTR rescued both cell growth and proliferation rate. {yields} MMTR blocked the CAK kinase-mediated phosphorylation of CDK1. {yields} The expression level of MMTR was modulated during cell cycle progression. -- Abstract: We have previously reported that MMTR (MAT1-mediated transcriptional repressor) is a co-repressor that inhibits TFIIH-mediated transcriptional activity via interaction with MAT1 (Kang et al., 2007). Since MAT1 is a member of the CAK kinase complex that is crucial for cell cycle progression and that regulates CDK phosphorylation as well as the general transcription factor TFIIH, we investigated MMTR function in cell cycle progression. We found that MMTR over-expression delayed G1/S and G2/M transitions, whereas co-expression of MAT1 and MMTR rescued the cell growth and proliferation rate. Moreover, MMTR was required for inhibition of CAK kinase-mediated CDK1 phosphorylation. We also showed that the expression level of MMTR was modulated during cell cycle progression. Our data support the notion that MMTR is an intrinsic negative cell cycle regulator that modulates the CAK kinase activity via interaction with MAT1.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2011

The thioredoxin TRX-1 regulates adult lifespan extension induced by dietary restriction in Caenorhabditis elegans

Highlights: {yields} First in vivo data for thioredoxin in dietary-restriction-(DR)-induced longevity. {yields} Thioredoxin (trx-1) loss suppresses longevity of eat-2 mutant, a genetic DR model. {yields} trx-1 overexpression extends wild-type longevity, but not that of eat-2 mutant. {yields} Longevity by dietary deprivation (DD), a non-genetic DR model, requires trx-1. {yields} trx-1 expression in ASJ neurons of aging adults is increased in response to DD. -- Abstract: Dietary restriction (DR) is the only environmental intervention known to extend adult lifespan in a wide variety of animal models. However, the genetic and cellular events that mediate the anti-aging programs induced by DR remain elusive. Here, we used the nematode Caenorhabditis elegans to provide the first in vivo evidence that a thioredoxin (TRX-1) regulates adult lifespan extension induced by DR. We found that deletion of the gene trx-1 completely suppressed the lifespan extension caused by mutation of eat-2, a genetic surrogate of DR in the worm. However, trx-1 deletion only partially suppressed the long lifespan caused by mutation of the insulin-like receptor gene daf-2 or by mutation of the sensory cilia gene osm-5. A trx-1::GFP translational fusion expressed from its own promoter in ASJ neurons (Ptrx-1::trx-1::GFP) rescued the trx-1 deletion-mediated suppression of the lifespan extension caused by mutation of eat-2. This rescue was not observed when trx-1::GFP was expressed from the ges-1 promoter in the intestine. In addition, overexpression of Ptrx-1::trx-1::GFP extended lifespan in wild type, but not in eat-2 mutants. trx-1 deletion almost completely suppressed the lifespan extension induced by dietary deprivation (DD), a non-genetic, nutrient-based model of DR in the worm. Moreover, DD upregulated the expression of a trx-1 promoter-driven GFP reporter gene (Ptrx-1::GFP) in ASJ neurons of aging adults, but not that of control Pgpa-9::GFP (which is also expressed in ASJ neurons). We propose that DR activates TRX-1 in ASJ neurons during aging, which in turn triggers TRX-1-dependent mechanisms to extend adult lifespan in the worm.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2011

Accelerated cellular senescence phenotype of GAPDH-depleted human lung carcinoma cells

Highlights: {yields} We examined the effect of glyceraldehyde 3-phosphate (GAPDH) depletion on proliferation of human carcinoma A549 cells. {yields} GAPDH depletion induces accelerated senescence in tumor cells via AMPK network, in the absence of DNA damage. {yields} Metabolic and genetic rescue experiments indicate that GAPDH has regulatory functions linking energy metabolism and cell cycle. {yields} Induction of senescence in LKB1-deficient lung cancer cells via GAPDH depletion suggests a novel strategy to control tumor cell proliferation. -- Abstract: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a pivotal glycolytic enzyme, and a signaling molecule which acts at the interface between stress factors and the cellular apoptotic machinery. Earlier, we found that knockdown of GAPDH in human carcinoma cell lines resulted in cell proliferation arrest and chemoresistance to S phase-specific cytotoxic agents. To elucidate the mechanism by which GAPDH depletion arrests cell proliferation, we examined the effect of GAPDH knockdown on human carcinoma cells A549. Our results show that GAPDH-depleted cells establish senescence phenotype, as revealed by proliferation arrest, changes in morphology, SA-{beta}-galactosidase staining, and more than 2-fold up-regulation of senescence-associated genes DEC1 and GLB1. Accelerated senescence following GAPDH depletion results from compromised glycolysis and energy crisis leading to the sustained AMPK activation via phosphorylation of {alpha} subunit at Thr172. Our findings demonstrate that GAPDH depletion switches human tumor cells to senescent phenotype via AMPK network, in the absence of DNA damage. Rescue experiments using metabolic and genetic models confirmed that GAPDH has important regulatory functions linking the energy metabolism and the cell cycle networks. Induction of senescence in LKB1-deficient non-small cell lung cancer cells via GAPDH depletion suggests a novel strategy to control tumor cell proliferation.

60 APPLIED LIFE SCIENCES↗