Phosphorylation by the stress-activated MAPK Slt2 down-regulates the yeast TOR complex 2

Saccharomyces cerevisiae target of rapamycin (TOR) complex 2 (TORC2) is an essential regulator of plasma membrane lipid and protein homeostasis. How TORC2 activity is modulated in response to changes in the status of the cell envelope is unclear. Here we document that TORC2 subunit Avo2 is a direct target of Slt2, the mitogen-activated protein kinase (MAPK) of the cell wall integrity pathway. Activation of Slt2 by overexpression of a constitutively active allele of an upstream Slt2 activator (Pkc1) or by auxin-induced degradation of a negative Slt2 regulator (Sln1) caused hyperphosphorylation of Avo2 at its MAPK phosphoacceptor sites in a Slt2-dependent manner and diminished TORC2-mediated phosphorylation of its major downstream effector, protein kinase Ypk1. Deletion of Avo2 or expression of a phosphomimetic Avo2 allele rendered cells sensitive to two stresses (myriocin treatment and elevated exogenous acetic acid) that the cell requires Ypk1 activation by TORC2 to survive. Thus, Avo2 is necessary for optimal TORC2 activity, and Slt2-mediated phosphorylation of Avo2 down-regulates TORC2 signaling. Compared with wild-type Avo2, phosphomimetic Avo2 shows significant displacement from the plasma membrane, suggesting that Slt2 inhibits TORC2 by promoting Avo2 dissociation. Our findings are the first demonstration that TORC2 function is regulated by MAPK-mediated phosphorylation.Comment: This work was supported by National Institutes of Health (NIH) Predoctoral Traineeship GM07232 and a University of California at Berkeley MacArthur and Lakhan-Pal Graduate Fellowship to K.L.L., Erwin Schroedinger Fellowship J3787-B21 from the Austrian Science Fund to AE-A, Marie Sklodowska-Curie Action H2020-MSCA-IF-2016 InsiliCardio, GA 75083 to CMA, and NIH R01 research grant GM21841 to J

Classical and all-floating FETI methods for the simulation of arterial tissues

High-resolution and anatomically realistic computer models of biological soft tissues play a significant role in the understanding of the function of cardiovascular components in health and disease. However, the computational effort to handle fine grids to resolve the geometries as well as sophisticated tissue models is very challenging. One possibility to derive a strongly scalable parallel solution algorithm is to consider finite element tearing and interconnecting (FETI) methods. In this study we propose and investigate the application of FETI methods to simulate the elastic behavior of biological soft tissues. As one particular example we choose the artery which is - as most other biological tissues - characterized by anisotropic and nonlinear material properties. We compare two specific approaches of FETI methods, classical and all-floating, and investigate the numerical behavior of different preconditioning techniques. In comparison to classical FETI, the all-floating approach has not only advantages concerning the implementation but in many cases also concerning the convergence of the global iterative solution method. This behavior is illustrated with numerical examples. We present results of linear elastic simulations to show convergence rates, as expected from the theory, and results from the more sophisticated nonlinear case where we apply a well-known anisotropic model to the realistic geometry of an artery. Although the FETI methods have a great applicability on artery simulations we will also discuss some limitations concerning the dependence on material parameters.Comment: 29 page

Roll convection of binary fluid mixtures in porous media

We investigate theoretically the nonlinear state of ideal straight rolls in the Rayleigh-B\'enard system of a fluid layer heated from below with a porous medium using a Galerkin method. Applying the Oberbeck-Boussinesq approximation, binary mixtures with positive separation ratio are studied and compared to one-component fluids. Our results for the structural properties of roll convection resemble qualitatively the situation in the Rayleigh--B\'enard system without porous medium except for the fact that the streamlines of binary mixtures are deformed in the so-called Soret regime. The deformation of the streamlines is explained by means of the Darcy equation which is used to describe the transport of momentum. In addition to the properties of the rolls, their stability against arbitrary infinitesimal perturbations is investigated. We compute stability balloons for the pure fluid case as well as for a wide parameter range of Lewis numbers and separation ratios which are typical for binary gas and fluid mixtures. The stability regions of rolls are found to be restricted by a crossroll, a zigzag and a new type of oscillatory instability mechanism, which can be related to the crossroll mechanism

Discovery and Metabolic Engineering of Steroid Alkaloid Biosynthetic Genes from (\u3ci\u3eVeratrum californicum\u3c/i\u3e)

The steroid alkaloid cyclopamine has shown much promise as a treatment for cancers in which aberrant hedgehog signaling plays a role. The compound, originally discovered due to its teratogenic effects in sheep, binds to the hedgehog signaling receptor Smoothend and prevents downstream activation. As this pathway is primarily active during embryonic development, overactivation later in life can lead to tumor formation and proliferation. Several studies have shown that cyclopamine can inhibit and even reverse tumor growth, but limited supply will prevent widespread use upon FDA approval of it or any of its semi-synthetic analogs. As high value plant medicinal compounds are in demand, producing them at industrially feasible levels is not always possible. Many plant natural products are originally produced in low quantities and often from species that are not amendable to cultivation. Production of these compounds in a heterologous system is ideal, but first the underlying genes that encode biosynthetic enzymes must be discovered. Cyclopamine is produced in one of these non-ideal sources, the slow growing Veratrum californicum, a species that has not been successfully cultivated. Our goal was to discover the underlying genes in the biosynthetic pathway to cyclopamine and express them in the emerging heterologous production system Camelina sativa. Herein, the first four genes in the biosynthetic steroid alkaloid pathway to verazine, a hypothesized intermediate of cyclopamine, have been discovered by correlating gene expression and alkaloid accumulation. In addition, the genes have been successfully introduced into C. sativa under the control of seed specific promoters for the production of V. californicum secondary metabolites in seed. Future work requires discovery of the remaining cyclopamine biosynthetic genes and metabolic fine tuning for increased metabolite yield, but the groundwork has been set for future work using C. sativa as a production system for high value, medicinally significant, plant natural products

Detecting small-scale spatial heterogeneity and temporal dynamics of soil organic carbon (SOC) stocks: a comparison between automatic chamber-derived C budgets and repeated soil inventories

Carbon (C) sequestration in soils plays a key role in the global C cycle. It is therefore crucial to adequately monitor dynamics in soil organic carbon (∆SOC) stocks when aiming to reveal underlying processes and potential drivers. However, small-scale spatial (10-30 m) and temporal changes in SOC stocks, particularly pronounced on arable lands, are hard to assess. The main reasons for this are limitations of the well-established methods. On the one hand, repeated soil inventories, often used in long-term field trials, reveal spatial patterns and trends in ∆SOC but require a longer observation period and a sufficient number of repetitions. On the other hand, eddy covariance measurements of C fluxes towards a complete C budget of the soil-plant-atmosphere system may help to obtain temporal ∆SOC patterns but lack small-scale spatial resolution. To overcome these limitations, this study presents a reliable method to detect both short-term temporal dynamics as well as small-scale spatial differences of ΔSOC using measurements of the net ecosystem carbon balance (NECB) as a proxy. To estimate the NECB, a combination of automatic chamber (AC) measurements of CO2 exchange and empirically modeled aboveground biomass development (NPPshoot) were used. To verify our method, results were compared with ΔSOC observed by soil resampling. Soil resampling and AC measurements were performed from 2010 to 2014 at a colluvial depression located in the hummocky ground moraine landscape of NE Germany. The measurement site is characterized by a variable groundwater level (GWL) and pronounced small-scale spatial heterogeneity regarding SOC and nitrogen (Nt) stocks. Tendencies and magnitude of ∆SOC values derived by AC-measurements and repeated soil inventories corresponded well. The period of maximum plant growth was identified as being most important for the development of spatial differences in annual ΔSOC. Hence, we were able to confirm that AC-based C budgets are able to reveal small-scale spatial differences and short-term temporal dynamics of ∆SOC

Effect of Tertiarybutylhydroquinone on the stability of fried banana chips

Perbandingan kestabilan terhadap pengoksidanan ke atas kerepek pisang yang digoreng dalam minyak kelapa sawit olein yang mengandungi antioksidan, tertiary"butylhydroquinone, dan minyak kelapa sawit olein tanpa antioksidan telah dikaji. Nilai-nilai peroksid d{m anisidin bagi minyak yang diekstrak dan kerepek yang disimpan pada suhu 60° C menjelaskan kerepek pisang yang digoreng dalam minyak mengandungi tertiarybu tylhydroquinone memperolehi kestabilan yang lebih baik terhadap jangkamasa penyimpanan bagi kerepek tersebu