The human silent information regulator (Sir)2 homologue hSIRT3 is a mitochondrial nicotinamide adenine dinucleotide-dependent deacetylase.

The yeast silent information regulator (Sir)2 protein links cellular metabolism and transcriptional silencing through its nicotinamide adenine dinucleotide (NAD)-dependent histone deacetylase activity. We report that mitochondria from mammalian cells contain intrinsic NAD-dependent deacetylase activity. This activity is inhibited by the NAD hydrolysis product nicotinamide, but not by trichostatin A, consistent with a class III deacetylase. We identify this deacetylase as the nuclear-encoded human Sir2 homologue hSIRT3, and show that hSIRT3 is located within the mitochondrial matrix. Mitochondrial import of hSIRT3 is dependent on an NH2-terminal amphipathic alpha-helix rich in basic residues. hSIRT3 is proteolytically processed in the mitochondrial matrix to a 28-kD product. This processing can be reconstituted in vitro with recombinant mitochondrial matrix processing peptidase (MPP) and is inhibited by mutation of arginines 99 and 100. The unprocessed form of hSIRT3 is enzymatically inactive and becomes fully activated in vitro after cleavage by MPP. These observations demonstrate the existence of a latent class III deacetylase that becomes catalytically activated upon import into the human mitochondria

Sirtuins: Sir2-related NAD-dependent protein deacetylases

Silent information regulator 2 (Sir2) proteins, or sirtuins, are protein deacetylases dependent on nicotine adenine dinucleotide (NAD) and are found in organisms ranging from bacteria to humans. In eukaryotes, sirtuins regulate transcriptional repression, recombination, the cell-division cycle, microtubule organization, and cellular responses to DNA-damaging agents. Sirtuins have also been implicated in regulating the molecular mechanisms of aging. The Sir2 catalytic domain, which is shared among all sirtuins, consists of two distinct domains that bind NAD and the acetyl-lysine substrate, respectively. In addition to the catalytic domain, eukaryotic sirtuins contain variable amino- and carboxy-terminal extensions that regulate their subcellular localizations and catalytic activity

Lifespan-increasing drug nordihydroguaiaretic acid inhibits p300 and activates autophagy.

Aging is characterized by the progressive loss of physiological function in all organisms. Remarkably, the aging process can be modulated by environmental modifications, including diet and small molecules. The natural compound nordihydroguaiaretic acid (NDGA) robustly increases lifespan in flies and mice, but its mechanism of action remains unclear. Here, we report that NDGA is an inhibitor of the epigenetic regulator p300. We find that NDGA inhibits p300 acetyltransferase activity in vitro and suppresses acetylation of a key p300 target in histones (i.e., H3K27) in cells. We use the cellular thermal shift assay to uniquely demonstrate NDGA binding to p300 in cells. Finally, in agreement with recent findings indicating that p300 is a potent blocker of autophagy, we show that NDGA treatment induces autophagy. These findings identify p300 as a target of NDGA and provide mechanistic insight into its role in longevity

Review of FEWS NET Biophysical Monitoring Requirements

The Famine Early Warning System Network (FEWS NET) provides monitoring and early warning support to decision makers responsible for responding to famine and food insecurity. FEWS NET transforms satellite remote sensing data into rainfall and vegetation information that can be used by these decision makers. The National Aeronautics and Space Administration has recently funded activities to enhance remote sensing inputs to FEWS NET. To elicit Earth observation requirements, a professional review questionnaire was disseminated to FEWS NET expert end-users: it focused upon operational requirements to determine additional useful remote sensing data and; subsequently, beneficial FEWS NET biophysical supplementary inputs. The review was completed by over 40 experts from around the world, enabling a robust set of professional perspectives to be gathered and analyzed rapidly. Reviewers were asked to evaluate the relative importance of environmental variables and spatio-temporal requirements for Earth science data products, in particular for rainfall and vegetation products. The results showed that spatio-temporal resolution requirements are complex and need to vary according to place, time, and hazard: that high resolution remote sensing products continue to be in demand, and that rainfall and vegetation products were valued as data that provide actionable food security information

Modeling rain-fed maize vulnerability to droughts using the standardized precipitation index from satellite estimated rainfall-Southern Malawi case study

During 1990s, disaster risk reduction emerged as a novel, proactive approach to managing risks from natural hazards. The World Bank, USAlD, and other international donor agencies began making efforts to mainstream disaster risk reduction in countries whose population and economies were heavily dependent on rain-fed agriculture. This approach has more significance in light of the increasing climatic hazard patterns and the climate scenarios projected for different hazard prone countries in the world. The Famine Early Warning System Network (FEWS NET) has been monitoring the food security issues in the sub-Saharan Africa, Asia and in Haiti. FEWS NET monitors the rainfall and moisture availability conditions with the help of NOAA RFE2 data for deriving food security status in Africa. This paper highlights the efforts in using satellite estimated rainfall inputs to develop drought vulnerability models in the drought prone areas in Malawi. The satellite RFE2 based SPI corresponding to the critical tasseling and silking phases (in the months of January, February, and March) were statistically regressed with drought-induced yield losses at the district level. The analysis has shown that the drought conditions in February and early March lead to most damage to maize yields in this region. The district-wise vulnerabilities to drought were up scaled to obtain a regional maize vulnerability model for southern Malawi. The results would help in establishing an early monitoring mechanism for drought impact assessment, give the decision makers additional time to assess seasonal outcomes, and identify potential food-related hazards in Malawi

An automatic multi-stepping approach to aircraft ice prediction

Flying an aircraft in icing conditions may seriously degrade its aerodynamical performance and threaten the flight safety. Over the years, new technologies and improved procedures have limited the potential risks caused by aircraft icing. Experimental studies being very expensive, numerous computer codes have been developed to simulate ice shapes and tackle the problem. Typically in these codes, a flow solution and key icing parameters are evaluated around a clean un-iced geometry and their values remain constant during the entire simulation. This approach may be acceptable for short exposure times or when the ice shape only slightly deforms the initial geometry. However, in other cases, the values of the icing parameters may vary and the simulation will loose its accuracy: for large shapes, the presence of the ice influences the surrounding airflow significantly, altering the value of icing parameters and ultimately the ice accretion. Calculating more accurate ice shapes therefore requires to periodically recompute the flow field around the body during the simulation and determine updated values for icing parameters. This procedure, known as multi-stepping, is investigated in this thesis and adapted to the new threedimensional icing code ICECREMO2. Several multi-step algorithms are presented and tested on cylinders and airfoils. When possible, the ice shapes simulated are compared with experimental results. The first multi-step calculations were generally performed manually. The user had to perform a rather tedious work and inappropriate instructions could lead to severe inaccuracies in the simulations. To avoid these difficulties, a fully automated procedure will be developed including all stages of a multi-step computation. This significantly reduces user interaction and the overall computing time. The present research work forms part of the ICECREMO2 project. ICECREMO2 is a three-dimensional ice accretion and water flow code developed collaboratively by Airbus UK, BAe Systems, Dunlop Aerospace, Rolls-Royce, GKN Westland Helicopters, QinetiQ and Cranfield University under the auspices of the UK Department of Trade and Industry. iEThOS - Electronic Theses Online ServiceGBUnited Kingdo

Acetylation of the HIV-1 Tat protein by p300 is important for its transcriptional activity.

AbstractThe human immunodeficiency virus 1 (HIV-1) Tat protein activates transcriptional elongation by recruiting the positive transcription elongation factor (pTEFb) complex to the TAR RNA element, which is located at the 5′ extremity of all viral transcripts [1–3]. Tat also associates in vitro and in vivo with the transcriptional coactivator p300/CBP [4–6]. This association has been proposed to recruit the histone acetyltransferase (HAT) activity of p300 to the integrated HIV-1 promoter. We have observed that the purified p300 HAT domain acetylates recombinant Tat proteins in vitro and that Tat is acetylated in vivo. The major targets of acetylation by p300 are lysine residues (Lys50 and Lys51) in the arginine-rich motif (ARM) used by Tat to bind RNA and for nuclear import. Mutation of these residues in full-length recombinant Tat blocked its acetylation in vitro. Furthermore, mutation of these lysine residues to arginine markedly decreased the synergistic activation of he HIV promoter by Tat and p300 or by Tat and cyclin T1. These results demonstrate that acetylation of Tat by p300/CBP is important for its transcriptional activation of the HIV promoter

Reduced Mobility of the Alternate Splicing Factor (Asf) through the Nucleoplasm and Steady State Speckle Compartments

Compartmentalization of the nucleus is now recognized as an important level of regulation influencing specific nuclear processes. The mechanism of factor organization and the movement of factors in nuclear space have not been fully determined. Splicing factors, for example, have been shown to move in a directed manner as large intact structures from sites of concentration to sites of active transcription, but splicing factors are also thought to exist in a freely diffusible state. In this study, we examined the movement of a splicing factor, ASF, green fluorescent fusion protein (ASF–GFP) using time-lapse microscopy and the technique fluorescence recovery after photobleaching (FRAP). We find that ASF–GFP moves at rates up to 100 times slower than free diffusion when it is associated with speckles and, surprisingly, also when it is dispersed in the nucleoplasm. The mobility of ASF is consistent with frequent but transient interactions with relatively immobile nuclear binding sites. This mobility is slightly increased in the presence of an RNA polymerase II transcription inhibitor and the ASF molecules further enrich in speckles. We propose that the nonrandom organization of splicing factors reflects spatial differences in the concentration of relatively immobile binding sites

Drought Monitoring and Assessment: Remote Sensing and Modeling Approaches for the Famine Early Warning Systems Network

Drought monitoring is an essential component of drought risk management. It is usually carried out using drought indices/indicators that are continuous functions of rainfall and other hydrometeorological variables. This chapter presents a few examples of how remote sensing and hydrologic modeling techniques are being used to generate a suite of drought monitoring indicators at dekadal (10-day), monthly, seasonal, and annual time scales for several selected regions around the world. Satellite-based rainfall estimates are being used to produce drought indicators such as standardized precipitation index, dryness indicators, and start of season analysis. The Normalized Difference Vegetation Index is being used to monitor vegetation condition. Several satellite data products are combined using agrohydrologic models to produce multiple short- and long-term indicators of droughts. All the data sets are being produced and updated in near-real time to provide information about the onset, progression, extent, and intensity of drought conditions. The data and products produced are available for download from the Famine Early Warning Systems Network (FEWS NET) data portal at http:// earlywarning.usgs.gov. The availability of timely information and products support the decision-making processes in drought-related hazard assessment, monitoring, and management with the FEWS NET. The drought-hazard monitoring approach perfected by the U.S. Geological Survey for FEWS NET through the integration of satellite data and hydrologic modeling can form the basis for similar decision support systems. Such systems can operationally produce reliable and useful regional information that is relevant for local, district-level decision making