A mutually inhibitory feedback loop between the 20S proteasome and its regulator, NQO1

NAD(P)H:quinone-oxidoreductase-1 (NQO1) is a cytosolic enzyme that catalyzes the reduction of various quinones using flavin adenine dinucleotide (FAD) as a cofactor. NQO1 has been also shown to rescue proteins containing intrinsically unstructured domains, such as p53 and p73, from degradation by the 20S proteasome through an unknown mechanism. Here, we studied the nature of interaction between NQO1 and the 20S proteasome. Our study revealed a double negative feedback loop between NQO1 and the 20S proteasome, whereby NQO1 prevents the proteolytic activity of the 20S proteasome and the 20S proteasome degrades the apo form of NQO1. Furthermore, we demonstrate, both in vivo and in vitro, that NQO1 levels are highly dependent on FAD concentration. These observations suggest a link between 20S proteolysis and the metabolic cellular state. More generally, the results may represent a regulatory mechanism by which associated cofactors dictate the stability of proteins, thus coordinating protein levels with the metabolic status.     

The interface of protein structure, protein biophysics, and molecular evolution

Abstract The interface of protein structural biology, protein biophysics, molecular evolution, and molecular population genetics forms the foundations for a mechanistic understanding of many aspects of protein biochemistry. Current efforts in interdisciplinary protein modeling are in their infancy and the state-of-the art of such models is described. Beyond the relationship between amino acid substitution and static protein structure, protein function, and corresponding organismal fitness, other considerations are also discussed. More complex mutational processes such as insertion and deletion and domain rearrangements and even circular permutations should be evaluated. The role of intrinsically disordered proteins is still controversial, but may be increasingly important to consider. Protein geometry and protein dynamics as a deviation from static considerations of protein structure are also important. Protein expression level is known to be a major determinant of evolutionary rate and several considerations including selection at the mRNA level and the role of interaction specificity are discussed. Lastly, the relationship between modeling and needed high-throughput experimental data as well as experimental examination of protein evolution using ancestral sequence resurrection and in vitro biochemistry are presented, towards an aim of ultimately generating better models for biological inference and prediction.     

Demographic and ecogeographic factors limit wild grapevine spread at the southern edge of its distribution range

The spatial distribution of plants is constrained by demographic and ecogeographic factors that determine the range and abundance of the species. Wild grapevine (Vitis vinifera ssp. sylvestris) is distributed from Switzerland in the north to Israel in the south. However, little is known about the ecogeographic constraints of this species and its genetic and phenotypic characteristics, especially at the southern edge of its distribution range in the Levant region. In this study, we explore the population structure of southern Levantine wild grapevines and the correlation between demographic and ecogeographic characteristics. Based on our genetic analysis, the wild grapevine populations in this region can be divided into two major subgroups in accordance with a multivariate spatial and ecogeographical clustering model. The identified subpopulations also differ in morphological traits, mainly leaf hairiness which may imply adaptation to environmental stress. The findings suggest that the Upper Jordan River population was spread to the Sea of Galilee area and that a third smaller subpopulation at the south of the Golan Heights may represent a distinguished gene pool or a recent establishment of a new population. A spatial distribution model indicated that distance to water sources, Normalized difference vegetation index, and precipitation are the main environmental factors constraining V. v. sylvestris distribution at its southern distribution range. These factors in addition to limited gene flow between populations prevent further spread of wild grapevines southwards to semi‐arid regions.     

Identification of DNA-binding Proteins Using Structural, Electrostatic and Evolutionary Features

DNA-binding proteins (DBPs) participate in various crucial processes in the life-cycle of the cells, and the identification and characterization of these proteins is of great importance. We present here a random forests classifier for identifying DBPs among proteins with known 3D structures. First, clusters of evolutionarily conserved regions (patches) on the surface of proteins were detected using the PatchFinder algorithm; earlier studies showed that these regions are typically the functionally important regions of proteins. Next, we trained a classifier using features like the electrostatic potential, cluster-based amino acid conservation patterns and the secondary structure content of the patches, as well as features of the whole protein, including its dipole moment. Using 10-fold cross-validation on a dataset of 138 DBPs and 110 proteins that do not bind DNA, the classifier achieved a sensitivity and a specificity of 0.90, which is overall better than the performance of published methods. Furthermore, when we tested five different methods on 11 new DBPs that did not appear in the original dataset, only our method annotated all correctly.The resulting classifier was applied to a collection of 757 proteins of known structure and unknown function. Of these proteins, 218 were predicted to bind DNA, and we anticipate that some of them interact with DNA using new structural motifs. The use of complementary computational tools supports the notion that at least some of them do bind DNA.     

Ruptured uterus in pregnancy: a Canadian hospital's experience.

Between 1966 and 1985, 15 cases of complete rupture of the uterus in pregnancy were identified among 52,854 deliveries at Foothills Provincial General Hospital, Calgary, for an incidence rate of 0.3 per 1000 deliveries. Previous cesarean section (in seven patients) was not the only predisposing factor: a history of dilatation and curettage (in two patients) or laparoscopy (in one) were also implicated. Long, obstructed labour did not appear to be a factor. Rupture also occurred in patients at low risk. The most frequent immediate complication was hypotension, in five patients. The rupture site was repaired in 11 of the patients; the other 4 underwent hysterectomy. Close surveillance and prompt intervention are the keys to good fetal and maternal outcome.     

Kinetics of cyclocreatine and Na+ cotransport in human breast cancer cells: mechanism of activity

The growth-inhibitory effect of cyclocreatine(CCr) and the kinetics of CCr andNa⁺ cotransport were investigatedin MCF7 human breast cancer cells and its adriamycin-resistant sublinewith use of ³¹P- and²³Na-NMR spectroscopy. Thegrowth-inhibitory effect in the resistant line occurred at a lower CCrconcentration and was more pronounced than in the wild-type line. Thiscorrelated with an ~10-fold higher affinity of CCr to the transporterin the resistant line. The passive diffusion coefficient of CCr wasalso higher in the resistant line by three- to fourfold. The transportof CCr was accompanied by a rapid increase in intracellularNa⁺. This increase was found todepend on the rate of CCr transport and varied differently with CCrconcentration in the two cell lines. It is proposed that thecotransport of CCr and Na⁺followed by increased Na⁺concentration, together with the accumulation of the highly charged phosphocyclocreatine, are responsible for cell swelling and death.