EC(5)S Ubiquitin Complex Is Recruited by KSHV Latent Antigen LANA for Degradation of the VHL and p53 Tumor Suppressors

Cellular protein degradation pathways can be utilized by viruses to establish an environment that favors their propagation. Here we report that the Kaposi's sarcoma–associated herpesvirus (KSHV)-encoded latency-associated nuclear antigen (LANA) directly functions as a component of the EC(5)S ubiquitin complex targeting the tumor suppressors von Hippel-Lindau (VHL) and p53 for degradation. We have characterized a suppressor of cytokine signaling box-like motif within LANA composed of an Elongin B and C box and a Cullin box, which is spatially located at its amino and carboxyl termini. This motif is necessary for LANA interaction with the Cul5–Elongin BC complex, to promote polyubiquitylation of cellular substrates VHL and p53 in vitro via its amino- and carboxyl-terminal binding domain, respectively. In transfected cells as well as KSHV-infected B lymphoma cells, LANA expression stimulates degradation of VHL and p53. Additionally, specific RNA interference–mediated LANA knockdown stabilized VHL and p53 in primary effusion lymphoma cells. Thus, manipulation of tumor suppressors by LANA potentially provides a favorable environment for progression of KSHV-infected tumor cells

Field-scale mapping of evaporative stress indicators of crop yield: An application over Mead, NE, USA

The Evaporative Stress Index (ESI) quantifies temporal anomalies in a normalized evapotranspiration (ET) metric describing the ratio of actual-to-reference ET (fRET) as derived from satellite remote sensing. At regional scales (3–10 km pixel resolution), the ESI has demonstrated the capacity to capture developing crop stress and impacts on regional yield variability in water-limited agricultural regions. However, its performance in some regions where the vegetation cycle is intensively managed appears to be degraded due to spatial and temporal limitations in the standard ESI products. In this study, we investigated potential improvements to ESI by generating maps of ET, fRET, and fRET anomalies at high spatiotemporal resolution (30-m pixels, daily time steps) using a multi-sensor data fusion method, enabling separation of landcover types with different phenologies and resilience to drought. The study was conducted for the period 2010–2014 covering a region around Mead, Nebraska that includes both rainfed and irrigated crops. Correlations between ESI and measurements of maize yield were investigated at both the field and county level to assess the potential of ESI as a yield forecasting tool. To examine the role of crop phenology in yield-ESI correlations, annual input fRET time series were aligned by both calendar day and by biophysically relevant dates (e.g. days since planting or emergence). At the resolution of the operational U.S. ESI product (4 km), adjusting fRET alignment to a regionally reported emergence date prior to anomaly computation improves r2 correlations with county-level yield estimates from 0.28 to 0.80. At 30-m resolution, where pure maize pixels can be isolated from other crops and landcover types, county-level yield correlations improved from 0.47 to 0.93 when aligning fRET by emergence date rather than calendar date. Peak correlations occurred 68 days after emergence, corresponding to the silking stage for maize when grain development is particularly sensitive to soil moisture deficiencies. The results of this study demonstrate the utility of remotely sensed ET in conveying spatially and temporally explicit water stress information to yield prediction and crop simulation models

Restriction landmark genomic scanning (RLGS) spot identification by second generation virtual RLGS in multiple genomes with multiple enzyme combinations.

BackgroundRestriction landmark genomic scanning (RLGS) is one of the most successfully applied methods for the identification of aberrant CpG island hypermethylation in cancer, as well as the identification of tissue specific methylation of CpG islands. However, a limitation to the utility of this method has been the ability to assign specific genomic sequences to RLGS spots, a process commonly referred to as "RLGS spot cloning."ResultsWe report the development of a virtual RLGS method (vRLGS) that allows for RLGS spot identification in any sequenced genome and with any enzyme combination. We report significant improvements in predicting DNA fragment migration patterns by incorporating sequence information into the migration models, and demonstrate a median Euclidian distance between actual and predicted spot migration of 0.18 centimeters for the most complex human RLGS pattern. We report the confirmed identification of 795 human and 530 mouse RLGS spots for the most commonly used enzyme combinations. We also developed a method to filter the virtual spots to reduce the number of extra spots seen on a virtual profile for both the mouse and human genomes. We demonstrate use of this filter to simplify spot cloning and to assist in the identification of spots exhibiting tissue-specific methylation.ConclusionThe new vRLGS system reported here is highly robust for the identification of novel RLGS spots. The migration models developed are not specific to the genome being studied or the enzyme combination being used, making this tool broadly applicable. The identification of hundreds of mouse and human RLGS spot loci confirms the strong bias of RLGS studies to focus on CpG islands and provides a valuable resource to rapidly study their methylation

Simultaneous determination of natural and synthetic steroid estrogens and their conjugates in aqueous matrices by liquid chromatography / mass spectrometry

An analytical method for the simultaneous determination of nine free and conjugated steroid estrogens was developed with application to environmental aqueous matrices. Solid phase extraction (SPE) was employed for isolation and concentration, with detection by liquid chromatography/mass spectrometry (LC/MS) using electrospray ionisation (ESI) in the negative mode. Method recoveries for various aqueous matrices (wastewater, lake and drinking water) were determined, recoveries proving to be sample dependent. When spiked at 50 ng/l concentrations in sewage influent, recoveries ranged from 62-89 % with relative standard deviations (RSD) < 8.1 %. In comparison, drinking water spiked at the same concentrations had recoveries between 82-100 % with an RSD < 5%. Ion suppression is a known phenomenon when using ESI; hence its impact on method recovery was elucidated for raw sewage. Both ion suppression from matrix interferences and the extraction procedure has bearing on the overall method recovery. Analysis of municipal raw sewage identified several of the analytes of interest at ng/l concentrations, estriol (E3) being the most abundant. Only one conjugate, estrone 3-sulphate (E1-3S) was observe

Phenological corrections to a field-scale, ET-based crop stress indicator: An application to yield forecasting across the U.S. Corn Belt

Soil moisture deficiency is a major factor in determining crop yields in water-limited agricultural production regions. Evapotranspiration (ET), which consists of crop water use through transpiration and water loss through direct soil evaporation, is a good indicator of soil moisture availability and vegetation health. ET therefore has been an integral part of many yield estimation efforts. The Evaporative Stress Index (ESI) is an ET-based crop stress indicator that describes temporal anomalies in a normalized evapotranspiration metric as derived from satellite remote sensing. ESI has demonstrated the capacity to explain regional yield variability in water-limited regions. However, its performance in some regions where the vegetation cycle is intensively managed appears to be degraded due to interannual phenological variability. This investigation selected three study sites across the U.S. Corn Belt – Mead, NE, Ames, IA and Champaign, IL – to investigate the potential operational value of 30-m resolution, phenologically corrected ESI datasets for yield prediction. The analysis was conducted over an 8-year period from 2010 to 2017, which included both drought and pluvial conditions as well as a broad range in yield values. Detrended yield anomalies for corn and soybean were correlated with ESI computed using annual ET curves temporally aligned based on (1) calendar date, (2) crop emergence date, and (3) a growing degree day (GDD) scaled time axis. Results showed that ESI has good correlations with yield anomalies at the county scale and that phenological corrections to the annual temporal alignment of the ET timeseries improve the correlation, especially when the time axis is defined by GDD rather than the calendar date. Peak correlations occur in the silking stage for corn and the reproductive stage for soybean – phases when these crops are particularly sensitive to soil moisture deficiencies. Regression equations derived at the time of peak correlation were used to estimate yields at county scale using a leave-one-out cross-validation strategy. The ESI-based yield estimates agree well with the USDA National Agricultural Statistics Service (NASS) county-level crop yield data, with correlation coefficients ranging from 0.79 to 0.93 and percent root-mean-square errors of 5–8%. These results demonstrate that remotely sensed ET at high spatiotemporal resolution can convey valuable water stress information for forecasting crop yields across the Corn Belt if interannual phenological variability is considered

A vertebrate case study of the quality of assemblies derived from next-generation sequences

The unparalleled efficiency of next-generation sequencing (NGS) has prompted widespread adoption, but significant problems remain in the use of NGS data for whole genome assembly. We explore the advantages and disadvantages of chicken genome assemblies generated using a variety of sequencing and assembly methodologies. NGS assemblies are equivalent in some ways to a Sanger-based assembly yet deficient in others. Nonetheless, these assemblies are sufficient for the identification of the majority of genes and can reveal novel sequences when compared to existing assembly references

TESS Discovery of a Transiting Super-Earth in the π\pi Mensae System

We report the detection of a transiting planet around $\pi$ Mensae (HD 39091), using data from the Transiting Exoplanet Survey Satellite (TESS). The solar-type host star is unusually bright (V=5.7) and was already known to host a Jovian planet on a highly eccentric, 5.7-year orbit. The newly discovered planet has a size of $2.04\pm 0.05$ $R_\oplus$ and an orbital period of 6.27 days. Radial-velocity data from the HARPS and AAT/UCLES archives also displays a 6.27-day periodicity, confirming the existence of the planet and leading to a mass determination of $4.82\pm 0.85$ $M_\oplus$. The star's proximity and brightness will facilitate further investigations, such as atmospheric spectroscopy, asteroseismology, the Rossiter--McLaughlin effect, astrometry, and direct imaging.Comment: Accepted for publication ApJ Letters. This letter makes use of the TESS Alert data, which is currently in a beta test phase. The discovery light curve is included in a table inside the arxiv submissio