Current studies on the Saccharomyces cerevisiae protein Dap1p have demonstrated a heme-related function within the ergosterol biosynthetic pathway. Here we present data to further the understanding of the role of heme in the proper biological functioning of Dap1p in cellular processes. First, we examined the role of Dap1p in stabilizing the P450 enzyme, Erg11p, a key regulatory protein in ergosterol biosynthesis. Our data indicate that the absence of Dap1p does not affect Erg11p mRNA, protein expression levels, or the protein degradation rates in S. Cerevisaie. Second, in order to probe the role of heme in the biological functioning of Dap1p, we measured ferric and ferrous heme binding affinities for Dap1p and the mutant Dap1pY138F, as well as equilibrium midpoint reduction potentials of the Fe(III)/Fe(II) couples. Our results show that both wild-type and mutant proteins bind heme in a 1:1 fashion, possessing tight ferric heme affinities, KD values of 400 pM and 200 nM, respectively, but exhibiting weak ferrous affinities, 2 and 10 microM, respectively. Additionally, the measured reduction potential of Dap1p, which was found to be -307 mV, is similar to that of other monotyrosinate hemoproteins. Although previous reports show the weaker affinity of Dap1pY138F for ferric heme lowers the production of ergosterol with respect to wild-type Dap1p in S. pombe, we find that Dap1pY138F expression is still sufficient to rescue the growth sensitivity of dap1Delta to fluconazole and methyl methanesulfonate in S. cerevisiae. Various interpretations of these results are discussed with respect to Dap1p function in the cell.

Water is essential to human health and economic development due to its utilization in sanitation, agriculture, and energy. Supplying water to an expanding world population requires simultaneous consideration of multiple societal sectors competing for limited resources. Water conservation, supply augmentation, distribution, and treatment of contaminants must work in concert to ensure water sustainability. Water is linked to other sectors, and the quantity and quality of water resources are changing. The efficient use of water in agriculture, the largest user of water worldwide, via drip irrigation is described as is the use of energy-intensive reverse osmosis to supplement freshwater supplies. Efforts to manage watersheds and model their responses to severe weather events are discussed along with efforts to improve the predictability of their function. The regional competition for water resources impacts both energy and water supply reliability, which requires that nations balance both for sustainable economic development. The use of water and energy in the US is described which provides a lens through which to both rethink the interrelationship of water and energy as well as evaluate technological developments. Advances in nanotechnology are highlighted as one emerging technology. These results underscore the multifaceted nature of water sustainability, its interrelationship to energy and economic development, and the need to develop, manage and regulate water systems in a concerted manner.