A PP2A-B55-Mediated Crosstalk between TORC1 and TORC2 Regulates the Differentiation Response in Fission Yeast

Extracellular cues regulate cell fate, and this is mainly achieved through the engagement of specific transcriptional programs. The TORC1 and TORC2 complexes mediate the integration of nutritional cues to cellular behavior, but their interplay is poorly understood. Here, we use fission yeast to investigate how phosphatase activity participates in this interplay during the switch from proliferation to sexual differentiation. We find that loss of PP2A-B55$^{Pab1}$ enhances the expression of differentiation-specific genes and leads to premature conjugation. $\textit{pab1}$ deletion brings about a transcriptional profile similar to TORC1 inactivation, and deletion of $\textit{pab1}$ overcomes the repression of differentiation genes in cells overexpressing TORC1. Importantly, we show that this effect is mediated by an increased TORC2-AKT (Gad8) signaling. Under nutrient-rich conditions, PP2A-B55$^{Pab1}$ dephosphorylates Gad8 Ser546, repressing its activity. Conversely, TORC1 inactivation upon starvation leads to the inactivation of PP2A-B55$^{Pab1}$ through the Greatwall-Endosulfin pathway. This results in the activation of Gad8 and the commitment to differentiation. Thus, PP2A-B55$^{Pab1}$ enables a crosstalk between the two TOR complexes that controls cell-fate decisions in response to nutrient availability.We thank Dominique Helmlinger and Sergio Moreno for sharing unpublished results and strains, and for stimulating discussion. We thank Janni Petersen, Ronit Weisman, Kazuhiro Shiozaki, Mitsuhiro Yanagida, and Hisao Masukata for strains, constructs, and antibodies. We thank Toni Hurtado and Beata Grallert for critical reading of the manuscript. This work was supported by NFR FRIMEDBIO grant 214049. M.P. is the recipient of a Kreftforeningen postdoctoral fellowship (grant 5843744). N.C. has received funding from the European Union Seventh Framework Programme (FP7-PEOPLE-2013-COFUND) under grant agreement 609020 - Scientia Fellows. J.M. was supported by BBSRC research grants BB/N007697/1 and BB/M021483/1

Post-glacial evolution of alpine environments in the western Mediterranean region : The Laguna Seca record

In an effort to understand how alpine environments from the western Mediterranean region responded to climate variations since the last glacial-interglacial transition, a detailed chronological control and sedimentological analysis, supported by magnetic susceptibility, total organic carbon and C/N data, were carried out on the sedimentary record of Laguna Seca (LS). This is a latitudinal and altitudinally (2259 masl) key alpine wetland site located in the easternmost area of the Sierra Nevada, southern Iberian Peninsula, where sediments accumulated during Heinrich Stadial 1, Bolling-Allerod (B-A) and the Younger Dryas (YD) previously unrecorded in alpine Sierra Nevada. Climate controlled sedimentation in LS and three coarse-grained and one fine-grained facies association are differentiated, which help us decipher the paleoenvironmental evolution of LS: (1) subaerial cohesionless debris flows during a paraglacial stage; (2) till or nival diamicton during a small glacier/nivation hollow stage; (3) massive mudstone by suspension settling of clays into standing water during a lacustrine stage; and (4) frost-shattering breccia deposited inside the lacustrine stage, probably during the YD, and linked to a periglacial substage. The development of a previously existing small glacial cirque during the Last Glacial Maximum (LGM) in the LS basin at an elevation between 2500 and 2300 m could be supported by the important availability of slope sediments glacially-conditioned such as debris flows, reworked by paraglacial slope processes during the first deglaciation stages, confirming previous studies of landforms in the catchment area and the LGM-Equilibrium Line Altitude estimation above 2400 masl in Sierra Nevada. Mean sediment accumulation rates in the LS sedimentary units (4.21 and 0.28 mm/yr during the paraglacial small glacier/nivation stage and the lacustrine stage, respectively) confirm that geomorphic activity accelerated just after glaciers retreated due to a slope adjustment and high availability of glacially conditioned sediments. An abrupt change in paleoenvironmental and paleoclimatic conditions occurred in LS at ~ 15.7 cal kyr BP. This change was probably due to an increase in temperature and precipitation in the western Mediterranean region during the B-A. At LS, this resulted in significant ice-melt, forming a deep-water lake in LS with important organic matter contribution until the end of the Early Holocene (except in the YD when the lake level probably dropped), but elsewhere a general glacier recession in the Sierra Nevada and an expansion of the Mediterranean forest in the southern Iberian Peninsula. Finally, the general long-term aridification that occurred during the Middle Holocene until the present in the western Mediterranean region triggered an important environmental change transforming LS into an ephemeral wetland with an increase in aquatic productivity.Peer reviewe

The Holocene Cedrus pollen record from Sierra Nevada (S Spain), a proxy for climate change in N Africa

Comprehending the effects of climate variability and disturbance on forested ecosystems is paramount to successfully managing forest environments under future climate scenarios (e.g., global warming, aridi-fication increase). Changes in fossil pollen abundance in sedimentary archives record past vegetation dynamics at regional scales, mainly related to climate changes and, in the last few millennia, to human impact. Pollen records can thus provide long databases with information on how the environment reacted to climate change before the historical record. In this study, we synthesized fossil pollen data from seven sites from the Sierra Nevada in southern Spain to investigate the response of forests in the western Mediterranean area to millennial-scale climate changes and to human impact during the Holocene. In particular, here we focused on Cedrus pollen abundances, which most-likely originated from Northern Africa and were carried to Sierra Nevada by wind. Cedrus pollen has received little attention in the Iberian Peninsula palynological records, for it occurs in low concentrations and has an African source, and thus this article explores the potential to reconstruct its past history and climate. Although Cedrus abundances are generally lower than 1% in the studied pollen samples, a comparison with North African (Moroccan) Cedrus pollen records shows similar trends at long- and short-term time-scales. Therefore, this record could be used as a proxy for changes in this forest species in North Africa. As observed in the Sierra Nevada synthetic record, the increasing trend of Cedrus pollen during the Middle and Late Holocene closely correlates with decreasing summer insolation. This would have produced overall cooler annual temperatures in Northern Africa (Middle Atlas and Rif Mountains) as well as lower summer evaporation, benefiting the growth of this cool-adapted montane tree species while increasing available moisture during the summer, which is critical for this water-demanding species. Millennial-scale variability also characterizes the Sierra Nevada Cedrus synthetic pollen record. Cedrus abundance oscillations co-vary with well-known millennial-scale climatic variability that controlled cedar abundance and altitudinal distribution in montane areas of N Africa. (C) 2020 Elsevier Ltd. All rights reserved.Peer reviewe

Hrk1 Plays Both Hog1-Dependent and -Independent Roles in Controlling Stress Response and Antifungal Drug Resistance in Cryptococcus neoformans

The HOG (High Osmolarity Glycerol response) pathway plays a central role in controlling stress response, ergosterol biosynthesis, virulence factor production, and differentiation of Cryptococcus neoformans, which causes fatal fungal meningoencephalitis. Recent transcriptome analysis of the HOG pathway discovered a Hog1-regulated gene (CNAG_00130.2), encoding a putative protein kinase orthologous to Rck1/2 in Saccharomyces cerevisiae and Srk1 in Schizosaccharomyces pombe. Its function is not known in C. neoformans. The present study functionally characterized the role of Hrk1 in C. neoformans. Northern blot analysis confirmed that HRK1 expression depends on the Hog1 MAPK. Similar to the hog1Δ mutant, the hrk1Δ mutant exhibited almost complete resistance to fludioxonil, which triggers glycerol biosynthesis via the HOG pathway. Supporting this, the hrk1Δ mutant showed reduced intracellular glycerol accumulation and swollen cell morphology in response to fludioxonil, further suggesting that Hrk1 works downstream of the HOG pathway. However, Hrk1 also appeared to have Hog1-independent functions. Mutation of HRK1 not only further increased osmosensitivity of the hog1Δ mutant, but also suppressed increased azole-resistance of the hog1Δ mutant in an Erg11-independent manner. Furthermore, unlike the hog1Δ mutant, Hrk1 was not involved in capsule biosynthesis. Hrk1 was slightly involved in melanin production but dispensable for virulence of C. neoformans. These findings suggest that Hrk1 plays both Hog1-dependent and –independent roles in stress and antifungal drug susceptibility and virulence factor production in C. neoformans. Particularly, the finding that inhibition of Hrk1 substantially increases azole drug susceptibility provides a novel strategy for combination antifungal therapy

Revisiting Generalized Chaplygin Gas as a Unified Dark Matter and Dark Energy Model

In this paper, we revisit generalized Chaplygin gas (GCG) model as a unified dark matter and dark energy model. The energy density of GCG model is given as $\rho_{GCG}/\rho_{GCG0}=[B_{s}+(1-B_{s})a^{-3(1+\alpha)}]^{1/(1+\alpha)}$, where $\alpha$ and $B_s$ are two model parameters which will be constrained by type Ia supernova as standard candles, baryon acoustic oscillation as standard rulers and the seventh year full WMAP data points. In this paper, we will not separate GCG into dark matter and dark energy parts any more as adopted in the literatures. By using Markov Chain Monte Carlo method, we find the result: $\alpha=0.00126_{- 0.00126- 0.00126}^{+ 0.000970+ 0.00268}$ and $B_s= 0.775_{- 0.0161- 0.0338}^{+ 0.0161+ 0.0307}$.Comment: 6 pages, 4 figure

Sedation Modulates Frontotemporal Predictive Coding Circuits and the Double Surprise Acceleration Effect

Two important theories in cognitive neuroscience are predictive coding (PC) and the global workspace (GW) theory. A key research task is to understand how these two theories relate to one another, and particularly, how the brain transitions from a predictive early state to the eventual engagement of a brain-scale state (the GW). To address this question, we present a source-localization of EEG responses evoked by the local-global task—an experimental paradigm that engages a predictive hierarchy, which encompasses the GW. The results of our source reconstruction suggest three phases of processing. The first phase involves the sensory (here auditory) regions of the superior temporal lobe and predicts sensory regularities over a short timeframe (as per the local effect). The third phase is brain-scale, involving inferior frontal, as well as inferior and superior parietal regions, consistent with a global neuronal workspace (GNW; as per the global effect). Crucially, our analysis suggests that there is an intermediate (second) phase, involving modulatory interactions between inferior frontal and superior temporal regions. Furthermore, sedation with propofol reduces modulatory interactions in the second phase. This selective effect is consistent with a PC explanation of sedation, with propofol acting on descending predictions of the precision of prediction errors; thereby constraining access to the GNW

Additional value of screening for minor genes and copy number variants in hypertrophic cardiomyopathy

Introduction: Hypertrophic cardiomyopathy (HCM) is the most prevalent inherited heart disease. Next-generation sequencing (NGS) is the preferred genetic test, but the diagnostic value of screening for minor and candidate genes, and the role of copy number variants (CNVs) deserves further evaluation. Methods: Three hundred and eighty-seven consecutive unrelated patients with HCM were screened for genetic variants in the 5 most frequent genes (MYBPC3, MYH7, TNNT2, TNNI3 and TPM1) using Sanger sequencing (N = 84) or NGS (N = 303). In the NGS cohort we analyzed 20 additional minor or candidate genes, and applied a proprietary bioinformatics algorithm for detecting CNVs. Additionally, the rate and classification of TTN variants in HCM were compared with 427 patients without structural heart disease. Results: The percentage of patients with pathogenic/likely pathogenic (P/LP) variants in the main genes was 33.3%, without significant differences between the Sanger sequencing and NGS cohorts. The screening for 20 additional genes revealed LP variants in ACTC1, MYL2, MYL3, TNNC1, GLA and PRKAG2 in 12 patients. This approach resulted in more inconclusive tests (36.0% vs. 9.6%, p<0.001), mostly due to variants of unknown significance (VUS) in TTN. The detection rate of rare variants in TTN was not significantly different to that found in the group of patients without structural heart disease. In the NGS cohort, 4 patients (1.3%) had pathogenic CNVs: 2 deletions in MYBPC3 and 2 deletions involving the complete coding region of PLN. Conclusions: A small percentage of HCM cases without point mutations in the 5 main genes are explained by P/LP variants in minor or candidate genes and CNVs. Screening for variants in TTN in HCM patients drastically increases the number of inconclusive tests, and shows a rate of VUS that is similar to patients without structural heart disease, suggesting that this gene should not be analyzed for clinical purposes in HCM

A Mathematical Model of Mitotic Exit in Budding Yeast: The Role of Polo Kinase

Cell cycle progression in eukaryotes is regulated by periodic activation and inactivation of a family of cyclin–dependent kinases (Cdk's). Entry into mitosis requires phosphorylation of many proteins targeted by mitotic Cdk, and exit from mitosis requires proteolysis of mitotic cyclins and dephosphorylation of their targeted proteins. Mitotic exit in budding yeast is known to involve the interplay of mitotic kinases (Cdk and Polo kinases) and phosphatases (Cdc55/PP2A and Cdc14), as well as the action of the anaphase promoting complex (APC) in degrading specific proteins in anaphase and telophase. To understand the intricacies of this mechanism, we propose a mathematical model for the molecular events during mitotic exit in budding yeast. The model captures the dynamics of this network in wild-type yeast cells and 110 mutant strains. The model clarifies the roles of Polo-like kinase (Cdc5) in the Cdc14 early anaphase release pathway and in the G-protein regulated mitotic exit network

Preclinical and randomized phase I studies of plitidepsin in adults hospitalized with COVID-19

Plitidepsin, a marine-derived cyclic-peptide, inhibits SARS-CoV-2 replication at nanomolar concentrations by targeting the host protein eukaryotic translation elongation factor 1A. Here, we show that plitidepsin distributes preferentially to lung over plasma, with similar potency against across several SARS-CoV-2 variants in preclinical studies. Simultaneously, in this randomized, parallel, open-label, proof-of-concept study (NCT04382066) conducted in 10 Spanish hospitals between May and November 2020, 46 adult hospitalized patients with confirmed SARS-CoV-2 infection received either 1.5 mg (n = 15), 2.0 mg (n = 16), or 2.5 mg (n = 15) plitidepsin once daily for 3 d. The primary objective was safety; viral load kinetics, mortality, need for increased respiratory support, and dose selection were secondary end points. One patient withdrew consent before starting procedures; 45 initiated treatment; one withdrew because of hypersensitivity. Two Grade 3 treatment-related adverse events were observed (hypersensitivity and diarrhea). Treatment-related adverse events affecting more than 5% of patients were nausea (42.2%), vomiting (15.6%), and diarrhea (6.7%). Mean viral load reductions from baseline were 1.35, 2.35, 3.25, and 3.85 log10 at days 4, 7, 15, and 31. Nonmechanical invasive ventilation was required in 8 of 44 evaluable patients (16.0%); six patients required intensive care support (13.6%), and three patients (6.7%) died (COVID-19-related). Plitidepsin has a favorable safety profile in patients with COVID-19

E2F1-Mediated Upregulation of p19INK4d Determines Its Periodic Expression during Cell Cycle and Regulates Cellular Proliferation

BACKGROUND: A central aspect of development and disease is the control of cell proliferation through regulation of the mitotic cycle. Cell cycle progression and directionality requires an appropriate balance of positive and negative regulators whose expression must fluctuate in a coordinated manner. p19INK4d, a member of the INK4 family of CDK inhibitors, has a unique feature that distinguishes it from the remaining INK4 and makes it a likely candidate for contributing to the directionality of the cell cycle. p19INK4d mRNA and protein levels accumulate periodically during the cell cycle under normal conditions, a feature reminiscent of cyclins. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we demonstrate that p19INK4d is transcriptionally regulated by E2F1 through two response elements present in the p19INK4d promoter. Ablation of this regulation reduced p19 levels and restricted its expression during the cell cycle, reflecting the contribution of a transcriptional effect of E2F1 on p19 periodicity. The induction of p19INK4d is delayed during the cell cycle compared to that of cyclin E, temporally separating the induction of these proliferative and antiproliferative target genes. Specific inhibition of the E2F1-p19INK4d pathway using triplex-forming oligonucleotides that block E2F1 binding on p19 promoter, stimulated cell proliferation and increased the fraction of cells in S phase. CONCLUSIONS/SIGNIFICANCE: The results described here support a model of normal cell cycle progression in which, following phosphorylation of pRb, free E2F induces cyclin E, among other target genes. Once cyclinE/CDK2 takes over as the cell cycle driving kinase activity, the induction of p19 mediated by E2F1 leads to inhibition of the CDK4,6-containing complexes, bringing the G1 phase to an end. This regulatory mechanism constitutes a new negative feedback loop that terminates the G1 phase proliferative signal, contributing to the proper coordination of the cell cycle and provides an additional mechanism to limit E2F activity