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This review summarizes knowledge concerning a ubiquitous plasma transmembrane protein family that mediates nucleobase or ascorbate secondary active transport (NAT). We show that prototype bacterial and mostly fungal members have become unique model systems to unravel structure-function relationships and regulation of expression, using classical and reverse genetics, as well as biochemical approaches. We discuss the importance of NAT-mediated ascorbate transport in mammals and how changes in substrate specificity, from different nucleobases to ascorbate, might have evolved at the molecular level. Finally, we also discuss how modelling NAT-purine interactions might constitute a step towards the use of NAT proteins as specific gateways for targeting pathogenic microbes.

Multivariate techniques and especially cluster analysis have been commonly used in Archaeometry. Exploratory and model-based techniques of clustering have been applied in geochemical (continuous) data of archaeological artifacts for provenance studies. Model-based clustering techniques like classification maximum-likelihood and mixture maximum likelihood had been used in a lesser extent in this context and although they seem to be suitable for such data, they either present practical difficulties -like high dimensionality of the data-or their performance give no evidence to support that they prevail on the standard methods (Papageorgiou et al., 2001). In this paper standard statistical methods (hierarchical clustering, principal components analysis) and the recently developed one of the multivariate mixture of normals with unknown number of components (see Dellaportas and Papageorgiou, 2005) in the category of the modelbased ones, are applied and compared. The data set comprises of chemical compositions in 188 ceramic samples derived from the Aegean islands and surrounding areas.

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In the UapA uric acid-xanthine permease of Aspergillus nidulans, subtle interactions between key residues of the putative substrate binding pocket, located in the TMS8-TMS9 loop (where TMS is transmembrane segment), and a specificity filter, implicating residues in TMS12 and the TMS1-TMS2 loop, are critical for function and specificity. By using a strain lacking all transporters involved in adenine uptake (DeltaazgA DeltafcyB DeltauapC) and carrying a mutation that partially inactivates the UapA specificity filter (F528S), we obtained 28 mutants capable of UapA-mediated growth on adenine. Seventy-two percent of mutants concern replacements of a single residue, R481, in the putative cytoplasmic loop TMS10-TMS11. Five missense mutations are located in TMS9, in TMS10 or in loops TMS1-TMS2 and TMS8-TMS9. Mutations in the latter loops concern residues previously shown to enlarge UapA specificity (Q113L) or to be part of a motif involved in substrate binding (F406Y). In all mutants, the ability of UapA to transport its physiological substrates remains intact, whereas the increased capacity for transport of adenine and other purines seems to be due to the elimination of elements that hinder the translocation of non-physiological substrates through UapA, rather than to an increase in relevant binding affinities. The additive effects of most novel mutations with F528S and allele-specific interactions of mutation R481G (TMS10-TMS11 loop) with Q113L (TMS1-TMS2 loop) or T526M (TMS12) establish specific interdomain synergy as a critical determinant for substrate selection. Our results strongly suggest that distinct domains at both sides of UapA act as selective dynamic gates controlling substrate access to their translocation pathway.