Mechanism of the Escherichia coli DNA T:G-mismatch endonuclease (Vsr protein) probed with thiophosphate-containing oligodeoxynucleotides

The mechanism of the Escherichia coli DNA T:G mismatch endonuclease (Vsr) has been investigated using oligodeoxynucleotides substituted, at the scissile phosphate, with isomeric phosphorothioates and a 3'-phosphorothiolate. Binding and kinetic data with the phosphorothioates/phosphorothiolate indicate that the two magnesium ions, which constitute essential co-factors, are required to stabilise the extra negative charge developed on the phosphate as the transition state is formed. Additionally one of the magnesium ions serves to activate the leaving group (the non-bridging 3'-oxygen atom of the scissile phosphate) during the hydrolysis reaction. Stereochemical analysis, using the R-p phosphorothioate isomer, indicates that Vsr carries out a hydrolytic reaction with inversion of stereochemistry at phosphorus, compatible with an in-line attack of water and a pentacovalent transition state with trigonal bipyramidal geometry. In conjunction with structures of Vsr bound to its products, these data allow the reconstruction of the enzyme-substrate complex and a comprehensive description of the hydrolysis mechanism. (c) 2005 Elsevier Ltd. All rights reserved.

Economic growth: a panacea for child labour?

In this paper, we test whether economic growth decreases child labour by bringing together data from the National Sample Survey of India and state-level macro data to estimate a bivariate probit model of schooling and labour. Our results lead us to conclude that contrary to popular wisdom, growth actually increases rather than decreases child labour because it increases the demand for child workers. The level of state NDP, village wages and household incomes are seen as the conduits through which growth influences the supply side of the child labour market.

A strategy for process-oriented validation of coupled chemistry–climate models

Evaluating CCMs with the presented framework will increase our confidence in predictions of stratospheric ozone change.