Regulation of the \u3cem\u3eEscherichia coli\u3c/em\u3e Tryptophan Operon by Early Reactions in the Aromatic Pathway

7-Methyltryptophan (7MT) or compounds which can be metabolized to 7MT, 3-methylanthranilic acid (3MA) and 7-methylindole, cause derepression of the trp operon through feedback inhibition of anthranilate synthetase. Tyrosine reverses 3MA or 7-methylindole derepression, apparently by increasing the amount of chorismic acid available to the tryptophan pathway. A mutant isolated on the basis of 3MA resistance (MAR 13) was found to excrete small amounts of chorismic acid and to have a feedback-resistant phenylalanine 3-deoxy-d-arabinoheptulosonic acid-7-phosphate (DAHP) synthetase. Genetic evidence indicates that the mutation conferring 3MA resistance and feedback resistance is very closely linked to aroG, the structural gene for the DAHP synthetase (phe). Since feedback inhibition of anthranilate synthetase by l-tryptophan (or 7MT) is competitive with chorismic acid, alterations in growth conditions (added tyrosine) or in a mutant (MAR 13) which increase the amount of chorismic acid available to the tryptophan pathway result in resistance to 7MT derepression. Owing to this competitive nature of tryptophan feedback inhibition of anthranilate synthetase by chorismic acid, the early pathway apparently serves to exert a regulatory influence on tryptophan biosynthesis

Mechanism of 3-Methylanthranilic Acid Derepression of the Tryptophan Operon in \u3cem\u3eEscherichia coli\u3c/em\u3e

3-Methylanthranilic acid (3MA) inhibits growth and causes derepression of the tryptophan biosynthetic enzymes in wild-type strains of Escherichia coli. Previous reports attributed this effect to an inhibition of the conversion of 1-(o-carboxyphenylamino)-1-deoxyribulose 5-phosphate to indole-3-glycerol phosphate and a consequent reduction in the concentration of endogenous tryptophan. Our studies have shown that 3MA-resistant mutants linked to the tryptophan operon have a feedback-resistant anthranilate synthetase; mutants with an altered indole-3-glycerol phosphate synthetase were not found. 3MA or 7-methylindole can be metabolized to 7-methyltryptophan, and 3MA, 7-methylindole, and 7-methyltryptophan lead to derepression of the tryptophan operon. Furthermore, 3MA-resistant mutants are also resistant to 7-methylindole derepression. These results strongly suggest that the primary cause of derepression by 3MA is through its conversion to 7-methyltryptophan, which can inhibit anthranilate synthetase, thereby decreasing the concentration of endogenous tryptophan. Unlike 5- or 6-methyltryptophan, 7-methyltryptophan does not appear to function as an active corepressor

Cosmid Cloning of Five \u3cem\u3eZymomonas trp\u3c/em\u3e Genes by Complementation of \u3cem\u3eEscherichia coli\u3c/em\u3e and \u3cem\u3ePseudomonas putida trp\u3c/em\u3e Mutants

A library of Zymomonas mobilis genomic DNA was constructed in the broad-host-range cosmid pLAFR1. The library was mobilized into a variety of Escherichia coli and Pseudomonas putida trp mutants by using the helper plasmid pRK2013. Five Z. mobilis trp genes were identified by the ability to complement the trp mutants. The trpF, trpB, and trpA genes were on one cosmid, while the trpD and trpC genes were on two separate cosmids. The organization of the Z. mobilis trp genes seems to be similar to the organization found in Rhizobium spp., Acinetobacter calcoaceticus, and Pseudomonas acidovorans. The trpF, trpB, and trpA genes appeared to be linked, but they were not closely associated with trpD or trpC genes

Effect of Tryptophan Analogs on Derepression of the \u3cem\u3eEscherichia coli\u3c/em\u3e Tryptophan Operon by Indole-3-Propionic Acid

The abilities of 14 tryptophan analogs to repress the tryptophan (trp) operon have been studied in Escherichia coli cells derepressed by incubation with 0.25 mM indole-3-propionic acid (IPA). trp operon expression was monitored by measuring the specific activities of anthranilate synthase (EC 4.1.3.27) and the tryptophan synthase (EC 4.2.1.20) β subunit. Analogs characterized by modification or removal of the α-amino group or the α-carboxyl group did not repress the trp operon. The only analogs among this group that appeared to interact with the trp aporepressor were IPA, which derepressed the trp operon, and d-tryptophan. Analogs with modifications of the indole ring repressed the trp operon to various degrees. 7-Methyl-tryptophan inhibited anthranilate synthase activity and consequently derepressed the trp operon. Additionally, 7-methyltryptophan prevented IPA-mediated derepression but, unlike tryptophan, did so in a non-coordinate manner, with the later enzymes of the operon being relatively more repressed than the early enzymes. The effect of 7-methyltryptophan on IPA-mediated derepression was likely not due to the interaction of IPA with the allosteric site of anthranilate synthase, even though feedback-resistant mutants of anthranilate synthase were partially resistant to derepression by IPA. The effect of 7-methyltryptophan on derepression by IPA was probably due to the effect of the analog-aporepressor complex on trp operon expression

A CFH12k lensing survey of X-ray luminous galaxy clusters. II. Weak lensing analysis and global correlations

Aims. We present a wide-field multi-color survey of a homogeneous sample of eleven clusters of galaxies for which we measure total masses and mass distributions from weak lensing. This sample, spanning a small range in both X-ray luminosity and redshift, is ideally suited to determining the normalisation of scaling relations between X-ray properties of clusters and their masses (the M − T_X and the M − L_X relations) and also estimating the scatter in these relations at a fixed luminosity. Methods. The eleven clusters in our sample are all X-ray luminous and span a narrow redshift range at z = 0.21 ± 0.04. The weak lensing analysis of the sample is based on ground-based wide-field imaging obtained with the CFH12k camera on CFHT. We use the methodology developed and applied previously on the massive cluster Abell 1689. A Bayesian method, implemented in the Im2shape software, is used to fit the shape parameters of the faint background galaxies and to correct for PSF smearing. A multi-color selection of the background galaxies is applied to retrieve the weak lensing signal, resulting in a background density of sources of ~10 galaxies per square arc minute. With the present data, shear profiles are measured in all clusters out to at least 2 Mpc (more than 15 from the center) with high confidence. The radial shear profiles are fitted with different parametric mass profiles and the virial mass M_(200) is estimated for each cluster and then compared to other physical properties. Results. Scaling relations between mass and optical luminosity indicate an increase of the M/L ratio with luminosity (M/L ∝ L^(0.8)) and a LX−M_(200) relation scaling as L_X ∝ M^(0.83±0.11)_(200) while the normalization of the M_(200) ∝ T^(3/2)_X relation is close to the one expected from hydrodynamical simulations of cluster formation as well as previous X-ray analyses. We suggest that the dispersion in the M_(200) − T_X and M_(200) − L_X relations reflects the different merging and dynamical histories for clusters of similar X-ray luminosities and intrinsic variations in their measured masses. Improved statistics of clusters over a wider mass range are required for a better control of the intrinsic scatter in scaling relations

‘Dominant ethnicity’ and the ‘ethnic-civic’ dichotomy in the work of A. D. Smith

This article considers the way in which the work of Anthony Smith has helped to structure debates surrounding the role of ethnicity in present-day nations. Two major lines of enquiry are evident here. First, the contemporary role of dominant ethnic groups within 'their' nations and second, the interplay between ethnic and civic elements in nationalist argument. The two processes are related, but maintain elements of distinctiveness. Smith's major contribution to the dominant ethnicity debate has been to disembed ethnicity from the ideologically-charged and/or anglo-centric discourse of ethnic relations and to place it in historical context, thereby opening up space for dominant group ethnicity to be considered as a distinct phenomenon. This said, Smith's work does not adequately account for the vicissitudes of dominant ethnicity in the contemporary West. Building on the classical works of Hans Kohn and Friedrich Meinecke, Anthony Smith has also made a seminal contribution to the debate on civic and ethnic forms of national identity and nationalist ideology. As well as freeing this debate from the strong normative overtones which it has often carried, he has continued to insist that the terms civic and ethnic should be treated as an ideal-typical distinction rather than a scheme of classification

Resting state correlates of subdimensions of anxious affect

Resting state fMRI may help identify markers of risk for affective disorder. Given the comorbidity of anxiety and depressive disorders and the heterogeneity of these disorders as defined by DSM, an important challenge is to identify alterations in resting state brain connectivity uniquely associated with distinct profiles of negative affect. The current study aimed to address this by identifying differences in brain connectivity specifically linked to cognitive and physiological profiles of anxiety, controlling for depressed affect. We adopted a two-stage multivariate approach. Hierarchical clustering was used to independently identify dimensions of negative affective style and resting state brain networks. Combining the clustering results, we examined individual differences in resting state connectivity uniquely associated with subdimensions of anxious affect, controlling for depressed affect. Physiological and cognitive subdimensions of anxious affect were identified. Physiological anxiety was associated with widespread alterations in insula connectivity, including decreased connectivity between insula subregions and between the insula and other medial frontal and subcortical networks. This is consistent with the insula facilitating communication between medial frontal and subcortical regions to enable control of physiological affective states. Meanwhile, increased connectivity within a frontoparietal-posterior cingulate cortex-precunous network was specifically associated with cognitive anxiety, potentially reflecting increased spontaneous negative cognition (e.g., worry). These findings suggest that physiological and cognitive anxiety comprise subdimensions of anxiety-related affect and reveal associated alterations in brain connectivity

Detection of Gravitational Lensing in the Cosmic Microwave Background

Gravitational lensing of the cosmic microwave background (CMB), a long-standing prediction of the standard cosmolgical model, is ultimately expected to be an important source of cosmological information, but first detection has not been achieved to date. We report a 3.4 sigma detection, by applying quadratic estimator techniques to all sky maps from the Wilkinson Microwave Anisotropy Probe (WMAP) satellite, and correlating the result with radio galaxy counts from the NRAO VLA Sky Survey (NVSS). We present our methodology including a detailed discussion of potential contaminants. Our error estimates include systematic uncertainties from density gradients in NVSS, beam effects in WMAP, Galactic microwave foregrounds, resolved and unresolved CMB point sources, and the thermal Sunyaev-Zeldovich effect.Comment: 27 pages, 20 figure