The 2'-5' RNA Ligase of Escherichia coli: Purification, Cloning, and Genomic Disruption

An RNA ligase previously detected in extracts of Escherichia coli is capable of joining Saccharomyces cerevisiae tRNA splicing intermediates in the absence of ATP to form a 2-5 phosphodiester linkage (Greer, C., Javor, B., and Abelson, J. (1983) Cell 33, 899-906). This enzyme specifically ligates tRNA half-molecules containing nucleoside base modifications and shows a preference among different tRNA species. In order to investigate the function of this enzyme in RNA metabolism, the ligase was purified to homogeneity from E. coli lysate utilizing chromatographic techniques and separation of proteins by SDS-polyacrylamide gel electrophoresis. A single polypeptide of approximately 20 kilodaltons exhibited RNA ligase activity. The amino terminus of this protein was sequenced, and the open reading frame (ORF) encoding it was identified by a data base search. This ORF, which encodes a novel protein with a predicted molecular mass of 19.9 kDa, was amplified from E. coli genomic DNA and cloned. ORFs coding for highly similar proteins were detected in Methanococcus jannaschii and Bacillus stearothermophilus. The chromosomal gene encoding RNA ligase in E. coli was disrupted, abolishing ligase activity in cell lysates. Cells lacking ligase activity grew normally under laboratory conditions. However, moderate overexpression of the ligase protein led to slower growth rates and a temperature-sensitive phenotype in both wild-type and RNA ligase knockout strains. The RNA ligase reaction was studied in vitro using purified enzyme and was found to be reversible, indicating that this enzyme may perform cleavage or ligation in vivo

Why bayesian “evidence for H1” in one condition and bayesian “evidence for H0” in another condition does not mean good-enough bayesian evidence for a difference between the conditions

Psychologists are often interested in whether an independent variable has a different effect in condition A than in condition B. To test such a question, one needs to directly compare the effect of that variable in the two conditions (i.e., test the interaction). Yet many researchers tend to stop when they find a significant test in one condition and a nonsignificant test in the other condition, deeming this as sufficient evidence for a difference between the two conditions. In this Tutorial, we aim to raise awareness of this inferential mistake when Bayes factors are used with conventional cutoffs to draw conclusions. For instance, some researchers might falsely conclude that there must be good-enough evidence for the interaction if they find good-enough Bayesian evidence for the alternative hypothesis, H1, in condition A and good-enough Bayesian evidence for the null hypothesis, H0, in condition B. The case study we introduce highlights that ignoring the test of the interaction can lead to unjustified conclusions and demonstrates that the principle that any assertion about the existence of an interaction necessitates the direct comparison of the conditions is as true for Bayesian as it is for frequentist statistics. We provide an R script of the analyses of the case study and a Shiny app that can be used with a 2 × 2 design to develop intuitions on this issue, and we introduce a rule of thumb with which one can estimate the sample size one might need to have a well-powered design

An engineered Tetrahymena tRNA(Gln) for in vivo incorporation of unnatural amino acids into proteins by nonsense suppression

A new tRNA, THG73, has been designed and evaluated as a vehicle for incorporating unnatural amino acids site-specifically into proteins expressed in vivo using the stop codon suppression technique. The construct is a modification of tRNAGln(CUA) from Tetrahymena thermophila, which naturally recognizes the stop codon UAG. Using electrophysiological studies of mutations at several sites of the nicotinic acetylcholine receptor, it is established that THG73 represents a major improvement over previous nonsense suppressors both in terms of efficiency and fidelity of unnatural amino acid incorporation. Compared with a previous tRNA used for in vivo suppression, THG73 is as much as 100-fold less likely to be acylated by endogenous synthetases of the Xenopus oocyte. This effectively eliminates a major concern of the in vivo suppression methodology, the undesirable incorporation of natural amino acids at the suppression site. In addition, THG73 is 4-10-fold more efficient at incorporating unnatural amino acids in the oocyte system. Taken together, these two advances should greatly expand the range of applicability of the in vivo nonsense suppression methodology

Factors Influencing Real Estate Brokerage Sales Staff Performance

This research examined factors that affect residential real estate sales staff performance from sixteen firms of various sizes in different residential markets. The human capital model was related to sales staff commission earnings. A psychological factors model and management systems model were introduced that explained variance beyond that accounted for by the human capital model. Multiple regression analysis demonstrated that five of the twenty-seven variables examined accounted for most of the statistically significant variance. These variables included gender, hours worked per week, amount of formal education, work satisfaction, and reputation of the firm. Suggestions on how to utilize the findings regarding those variables related to earnings, as well as those not related, are briefly discussed.

Cognitive consequences of perceiving social exclusion

Although a great deal is now known about how people mentally represent individuals and groups, less attention has been paid to the question of how interpersonal relationships are represented in memory. Drawing on principles of categorization, this paper reports an investigation into how we mentally represent the relationships of others. In three experiments, evidence for assimilation effects following social exclusion (and subsequent categorization) is found. Experiment 1 uses a judgment paradigm to demonstrate that social exclusion influences the perception of interpersonal closeness. Experiments 2 and 3 employ a memory confusion paradigm to establish that representations of relationship partners are assimilated following the exclusion of a third party. © 2008 Elsevier Inc. All rights reserved

Tourist choice processing: evaluating decision rules and methods of their measurement

A detailed understanding of decision rules is essential in order to better explain consumption behavior, yet the variety of decision rules used have been somewhat neglected in tourism research. This study adopts an innovative method, greedoid analysis, to estimate a noncompensatory type of decision rule known as lexicographic by aspect (LBA). It is quite different from the weighted additive (WADD) model commonly assumed in tourism studies. By utilizing an experimental research design, this study enables the evaluation of the two types of decision rules regarding their predictive and explanatory power. Additionally, we introduce a novel evaluation indicator (“cost”), which allows further investigation of the heterogeneity in the use of decision rules. The results suggest that although the out-of-sample accuracy is lower, the LBA model has a better explanatory performance on respondents’ preference order. Moreover, the different perspective provided by the LBA model is useful for obtaining managerial implications

Setting Priorities for Space Research: Opportunities and Imperatives

This report represents the first phase of a study by a task group convened by the Space Studies Board to ascertain whether it should attempt to develop a methodology for recommending priorities among the various initiatives in space research (that is, scientific activities concerned with phenomena in space or utilizing observations from space). The report argues that such priority statements by the space research community are both necessary and desirable and would contribute to the formulation and implementation of public policy. The report advocates the establishment of priorities to enhance effective management of the nation's scientific research program in space. It argues that scientific objectives and purposes should determine how and under what circumstances scientific research should be done. The report does not take a position on the controversy between advocates of manned space exploration and those who favor the exclusive use of unmanned space vehicles. Nor does the report address questions about the value or appropriateness of Space Station Freedom or proposals to establish a permanent manned Moon base or to undertake a manned mission to Mars. These issues lie beyond the charge to the task group

Biological And Symptom Changes In Posttraumatic Stress Disorder Treatment: A Randomized Clinical Trial

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110764/1/da22331.pd

Polarization of coalitions in an agent-based model of political discourse

Political discourse is the verbal interaction between political actors in a policy domain. This article explains the formation of polarized advocacy or discourse coalitions in this complex phenomenon by presenting a dynamic, stochastic, and discrete agent-based model based on graph theory and local optimization. In a series of thought experiments, actors compute their utility of contributing a specific statement to the discourse by following ideological criteria, preferential attachment, agenda-setting strategies, governmental coherence, or other mechanisms. The evolving macro-level discourse is represented as a dynamic network and evaluated against arguments from the literature on the policy process. A simple combination of four theoretical mechanisms is already able to produce artificial policy debates with theoretically plausible properties. Any sufficiently realistic configuration must entail innovative and path-dependent elements as well as a blend of exogenous preferences and endogenous opinion formation mechanisms

Channelized melt flow in downwelling mantle: Implications for 226Ra-210Pb disequilibria in arc magmas

We present the results of an analytical model of porous flow of viscous melt into a steadily dilating ‘‘channel’’ (defined as a cluster of smaller veins) in downwelling subarc mantle. The model predicts the pressure drop in the mantle wedge matrix surrounding the channel needed to drive melt flow as a function of position and time. Melt is sucked toward the dilatant region at a near-constant velocity (105 s1) until veins comprising the channel stop opening (t = t). Fluid elements that complete their journey within the time span t < t arrive at a channel. Our results make it possible to calculate the region of influence sampled by melt that surrounds the channel. This region is large compared to the model size of the channelized region driving flow. For a baseline dilation time of 1 year and channel half width of 2 m, melt can be sampled over an 80-m radius and has the opportunity to sample matrix material with potentially contrasting chemistry on geologically short timescales. Our mechanical results are consistent with a downgoing arc mantle wedge source region where melting and melt extraction by porous flow to a channel network are sufficiently rapid to preserve source-derived 238U-230Th-226Ra, and potentially also 226 Ra-210Pb, disequilibria, prior to magma ascent to the surface. Since this is the rate-determining step in the overall process, it allows the possibility that such short-lived disequilibria measured in arc rocks at the surface are derived from deep in the mantle wedge. Stresses due to partial melting do not appear capable of producing the desired sucking effect, while the order of magnitude rate of shear required to drive dilation of 107 s1 is much larger than values resulting from steady state subduction. We conclude that local deformation rates in excess of background plate tectonic rates are needed to ‘‘switch on’’ the dilatant channel network and to initiate the sucking effect