Genetic and Molecular Dissection of Arsenic Hyperaccumulation

We have constructed cDNA libraries from RNA isolated from arsenic treated gametophytes of the fern Pteris vittata. This library was made in a manner that allows each cDNA clone to be expressed in yeast. We have introduced this library into yeast cells, both wild type and arsensic sensitive mutants, and selected transformed yeast colonies with increased arsenic tolerance compared to the parental strains. From these screens we have identified putative homologs of the yeast ACR2 and ACR3 genes from Pteris vittata and, for the past year, have focused on characterizing the function of the ACR2 gene. In yeast, ACR2 is an arsenate reductase that is essential for arsenate tolerance. We refer to the Pteris vittata ACR2 gene as PvACR2. The deduced amino acid sequence of PvACR2 is highly similar to the yeast ACR2 and other related phosphatases