Structure of the Human TELO2-TTI1-TTI2 Complex

Phosphatidylinositol 3-kinase-related protein kinases (PIKKs) play critical roles in various metabolic pathways related to cell proliferation and survival. The TELO2-TTI1-TTI2 (TTT) complex has been proposed to recognize newly synthesized PIKKs and to deliver them to the R2TP complex (RUVBL1-RUVBL2-RPAP3-PIH1D1) and the heat shock protein 90 chaperone, thereby supporting their folding and assembly. Here, we determined the cryo-EM structure of the TTT complex at an average resolution of 4.2 Å. We describe the full-length structures of TTI1 and TELO2, and a partial structure of TTI2. All three proteins form elongated helical repeat structures. TTI1 provides a platform on which TELO2 and TTI2 bind to its central region and C-terminal end, respectively. The TELO2 C-terminal domain (CTD) is required for the interaction with TTI1 and recruitment of Ataxia-telangiectasia mutated (ATM). The N- and C-terminal segments of TTI1 recognize the FRAP-ATM-TRRAP (FAT) domain and the N-terminal HEAT repeats of ATM, respectively. The TELO2 CTD and TTI1 N- and C-terminal segments are required for cell survival in response to ionizing radiation.     

Synergistic interactions between leaf beetle herbivory and fire enhance tamarisk (Tamarix spp.) mortality

The combined effects of herbivory and fire on plant mortality were investigated using prescribed burns of tamarisk (Tamarix ramosissima Lebed) exposed to herbivory by the saltcedar leaf beetle (Chrysomelidae: Diorhabda carinulata Desbrocher). Tamarix stands in the Humboldt Sink (NV, USA) were divided into three treatments: summer burn (August 2006), fall burn (October 2006) and control (unburned), and litter depth was manipulated to vary fire intensity within burn seasons. A gradient of existing herbivory impact was described with three plant condition metrics prior to fire: reduced proportions of green canopy, percent root crown starch sampled at the height of the growing season (August 2006), and percent root crown starch measured during dormancy (December 2006). August root crown starch concentration and proportion green canopy were strongly correlated, although the proportion green canopy predicted mortality better than August root crown starch. December root crown starch concentration was more depleted in unburned trees and in trees burned during the summer than in fall burn trees. Mortality in summer burned trees was higher than fall burned trees due to higher fire intensity, but December root crown starch available for resprouting in the spring was also lower in summer burned trees. The greatest mortality was observed in trees with the lowest December root crown starch concentration which were exposed to high fire intensity. Disproportionate changes in the slope and curvature of prediction traces as fire intensity and December starch reach reciprocal maximum and minimum levels indicate that beetle herbivory and fire intensity are synergistic.     

CircNr1h4 regulates the pathological process of renal injury in salt‐sensitive hypertensive mice by targeting miR‐155‐5p

Circular RNAs are a class of widespread and diverse endogenous RNAs that may regulate gene expression in various diseases, but their regulation and function in hypertensive renal injury remain unclear. In this study, we generated ribosomal‐depleted RNA sequencing data from normal mouse kidneys and from injured mouse kidneys induced by deoxycorticosterone acetate‐salt hypertension and identified at least 4900 circRNA candidates. A total of 124 of these circRNAs were differentially expressed between the normal and injured kidneys. Furthermore, we characterized one abundant circRNA, termed circNr1h4, which is derived from the Nr1h4 gene and significantly down‐regulated in the injured kidneys. RNA sequencing data and qPCR analysis also showed many microRNAs and mRNAs, including miR‐155‐5p and fatty acid reductase 1 (Far1), were differentially expressed between the normal and injured kidney and related to circNr1h4. In vitro, the silencing of circNr1h4 or overexpression of miR‐155‐5p significantly decreased Far1 levels and increased reactive oxygen species. Mechanistic investigations indicated that circNr1h4 acts as a competing endogenous RNA for miR‐155‐5p, leading to regulation of its target gene Far1. Our study provides novel insight into the molecular mechanisms underlying kidney injury in hypertension, which will be required to develop therapeutic strategies of targeting circRNAs for hypertensive kidney injury.     

High-density lipoproteins (HDL): Novel function and therapeutic applications

The failure of high-density lipoprotein (HDL)-raising agents to reduce cardiovascular disease (CVD) together with recent findings of increased cardiovascular mortality in subjects with extremely high HDL-cholesterol levels provide new opportunities to revisit our view of HDL. The concept of HDL function developed to explain these contradictory findings has recently been expanded by a role played by HDL in the lipolysis of triglyceride-rich lipoproteins (TGRLs) by lipoprotein lipase. According to the reverse remnant-cholesterol transport (RRT) hypothesis, HDL critically contributes to TGRL lipolysis via acquirement of surface lipids, including free cholesterol, released from TGRL. Ensuing cholesterol transport to the liver with excretion into the bile may reduce cholesterol influx in the arterial wall by accelerating removal from circulation of atherogenic, cholesterol-rich TGRL remnants. Such novel function of HDL opens wide therapeutic applications to reduce CVD in statin-treated patients, which primarily involve activation of cholesterol flux upon lipolysis.     

Rat brain myo-inositol 3-phosphate synthase is a phosphoprotein

The therapeutic effects of lithium in bipolar disorder are poorly understood. Lithium decreases free inositol levels by inhibiting inositol monophosphatase 1 and myo-inositol 3-phosphate synthase (IPS). In this study, we demonstrate for the first time that IPS can be phosphorylated. This was evident when purified rat IPS was dephosphorylated by lambda protein phosphatase and analyzed by phospho-specific ProQ-Diamond staining and Western blot analysis. These techniques demonstrated a mobility shift consistent with IPS being phosphorylated. Mass spectral analysis revealed that Serine-524 (S₅₂₄), which resides in the hinge region derived from exon 11 of the gene, is the site for phosphorylation. Further, an antibody generated against a synthetic peptide of IPS containing monophosphorylated-S₅₂₄, was able to discriminate the phosphorylated and non-phosphorylated forms of IPS. The phosphoprotein is found in the brain and testis, but not in the intestine. The intestinal IPS isoform lacks the peptide bearing S₅₂₄, and hence, cannot be phosphorylated. Evidences suggest that IPS is monophosphorylated at S₅₂₄ and that the removal of this phosphate does not alter its enzymatic activity. These observations suggest a novel function for IPS in brain and other tissues. Future studies should resolve the functional role of phospho-IPS in brain inositol signaling.     

Electron paramagnetic resonance spectroscopy on G-protein-coupled receptors: Adopting strategies from related model systems

Membrane proteins, including ion channels, transporters and G-protein-coupled receptors (GPCRs), play a significant role in various physiological processes. Many of these proteins are difficult to express in large quantities, imposing crucial experimental restrictions. Nevertheless, there is now a wide variety of studies available utilizing electron paramagnetic resonance (EPR) spectroscopic techniques that expand experimental accessibility by using relatively small quantities of protein. Here, we give an overview starting from basic strategies in EPR on membrane proteins with a focus on GPCRs, while emphasizing several applications from recent years. We highlight how the arsenal of EPR-based techniques may provide significant further contributions to understanding the complex molecular machinery and energetic phenomena responsible for seamless workflow in essential biological processes.     

Indicators of seabird reproductive performance demonstrate the impact of commercial fisheries on seabird populations in the North Sea

In a world of growing anthropogenic pressures on biodiversity, effective indicators need to be specific and sensitive to the pressures in the ecosystem concerned, yet be simple enough to be interpreted by non-experts and straightforward enough to facilitate routine monitoring. Globally, seabirds are under increasing pressure as a result of anthropogenic activities and environmental variation. Traditionally, seabird indicators have been based on abundance at breeding colonies. However, as many species do not reach sexual maturity for several years, and may not attend the colony over this time period, such indicators may fail to capture the ecological complexity of the system concerned.We constructed two indicators of the state of nine seabird species that breed along the UK coast of the North Sea: (i) abundance of seabirds at breeding colonies, and (ii) probability of seabird breeding failure. The indicators were significantly and strongly correlated with each other for eight out of nine species, but the abundance indicator typically lagged the indicator on seabird breeding failure by two to three years. We then considered a third indicator which compared kittiwake (Rissa tridactyla) breeding success to the levels expected given the underlying environmental conditions; changes in the abundance indicator also lagged this by three years. We investigate how sensitive each of these indicators was to the impacts of fishing. We found that the species which had seen the greatest increases in breeding failure rate over the study period were those species which were most sensitive to fisheries pressure.By focussing on demographic parameters, and correcting for the underlying environmental conditions, we can detect potentially important population level changes at an earlier stage than by focussing on abundance alone. These indicators are able to more accurately capture the complexity of the ecosystem concerned and can be readily interpreted by policy-makers.     

C-type natriuretic peptide: A new cardiac mediator

Natriuretic peptides are endogenous hormones released by the heart in response to myocardial stretch and overload. While atrial and brain natriuretic peptides (ANP, BNP) were immediately considered cardiac hormones and their role was well-characterized and defined in predicting risk in cardiovascular disease, evidence indicating the role of C-type natriuretic peptide (CNP) in cardiovascular regulation was slow to emerge until about 8 years ago. Since then, considerable literature on CNP and the cardiovascular system has been published; the aim of this review is to examine current literature relating to CNP and cardiovascular disease, in particular its role in heart failure (HF) and myocardial infarction (MI). This review retraces the fundamental steps in research that led understanding the role of CNP in HF and MI; from increased CNP mRNA expression and plasmatic concentrations in humans and in animal models, to detection of CNP expression in cardiomyocytes, to its evaluation in human leukocytes. The traditional view of CNP as an endothelial peptide has been surpassed by the results of many studies published in recent years, and while its physiological role is still under investigation, information is now available regarding its contribution to cardiovascular function. Taken together, these observations suggest that CNP and its specific receptor, NPR-B, can play a very important role in regulating cardiac hypertrophy and remodeling, indicating NPR-B as a new potential drug target for the treatment of cardiovascular disease.