Molecular architecture of fur binding to iron-induced and - repressed genes in Helicobacter pylori

The ferric uptake regulator protein Fur regulates iron-dependent gene expression in bacteria. In the human pathogen Helicobacter pylori, Fur has been shown to regulate iron-induced and iron-repressed genes. Herein we investigate the molecular mechanisms that control this differential iron-responsive Fur regulation. Hydroxyl radical footprinting showed that Fur has different binding architectures, which characterize distinct operator typologies. On operators recognized with higher affinity by holo-Fur, the protein binds to a continuous AT-rich stretch of about 20 bp, displaying an extended protection pattern. This is indicative of protein wrapping around the DNA helix. DNA binding interference assays with the minor groove binding drug distamycin A, point out that the recognition of the holo-operators occurs through the minor groove of the DNA. By contrast, on the apo-operators, Fur binds primarily to thymine dimers within a newly identified TCATTn10TT consensus element, indicative of Fur binding to one side of the DNA, in the major groove of the double helix. Reconstitution of the TCATTn10TT motif within a holo-operator results in a feature binding swap from an holo-Fur- to an apo-Fur-recognized operator, affecting both affinity and binding architecture of Fur, and conferring apo-Fur repression features in vivo. Size exclusion chromatography indicated that Fur is a dimer in solution. However, in the presence of divalent metal ions the protein is able to multimerize. Accordingly, apo-Fur binds DNA as a dimer in gel shift assays, while in presence of iron, higher order complexes are formed. Stoichiometric Ferguson analysis indicates that these complexes correspond to one or two Fur tetramers, each bound to an operator element. Together these data suggest that the apo- and holo-Fur repression mechanisms apparently rely on two distinctive modes of operator-recognition, involving respectively the readout of a specific nucleotide consensus motif in the major groove for apo-operators, and the recognition of AT-rich stretches in the minor groove for holo-operators, whereas the iron-responsive binding affinity is controlled through metal-dependent shaping of the protein structure in order to match preferentially the major or the minor groove.

The Efficiency of Microcredit Applied to Social Exclusion.

Microcredit has been a tool to alleviate poverty since long. This research is aimed to observe the efficiency of microcredit in the field of social exclusion. The development of questionnaires and use of existing tools was used to observe the tangible and intangible intertwining of microcredit and by doing so the effort was concentrated to observe whether microcredit has a direct effect on social exclusion or not. Bangladesh was chosen for the field study and 85 samples were taken for the analysis. It is a time period research and one year time was set to receive the sample and working on the statistical analysis. The tangible aspect was based on a World Bank questionnaire and the social capital questionnaire was developed through different well observed tools. The borrowers of Grameen Bank in Bangladesh, is the research sample whish shows a strong correlation between their tangible activity and social life. There are significant changes in tangible aspect and social participation observed from the research. Strong correlation between the two aspects was also found taking into account that the borrowers themselves have a vibrant social life in the village.