Panel 3: ACOs in Practice: Research on Current Implementation of ACOs

Lessons from the Wisconsin ACO Study State-Based ACO and Medical Home Pilots: Early Lessons from the Other Washington Lessons from ACO Implementation in New Jersey Accountable Care Organizations: A New Thing with Some Old Problem

Panel 3: ACOs in Practice: Research on Current Implementation of ACOs

Lessons from the Wisconsin ACO Study State-Based ACO and Medical Home Pilots: Early Lessons from the Other Washington Lessons from ACO Implementation in New Jersey Accountable Care Organizations: A New Thing with Some Old Problem

Data from: Identification and evaluation of novel acetolactate synthase inhibitors as antifungal agents

High-throughput phenotypic screening against yeast Saccharomyces cerevisiae revealed a series of triazolo-pyrimidine-sulfonamide compounds with broad-spectrum antifungal activity, no significant cytotoxicity, and low protein binding. To elucidate the target of this series we have applied a chemogenomic profiling approach using the S. cerevisiae deletion collection. All compounds of the series yielded highly similar profiles that suggested acetolactate synthase (Ilv2p, catalyzes the first common step in branched chain amino acid biosynthesis) as a possible target. High correlation to profiles of known Ilv2p inhibitors like chlorimuron-ethyl provided further evidence for a similar mechanism of action. Genome-wide mutagenesis in S. cerevisiae identified 13 resistant clones with 3 different mutations in the catalytic subunit of acetolactate synthase that also conferred cross-resistance to established Ilv2p inhibitors. Mapping the mutations into the published Ilv2p crystal structure outlined the chlorimuron-ethyl binding cavity and it was possible to dock the triazolo-pyrimidine-sulfonamide compound into this pocket in silico. However, fungal growth inhibition could be bypassed through supplementation with exogenous branched chain amino acids, or by the addition of serum to the medium in all of the fungal organisms tested except for Aspergillus fumigatus. Thus, these data support the identification of triazolo-pyrimidine-sulfonamide as inhibitors of acetolactate synthase but suggest that targeting may be compromised due to the possibility of nutrient bypass in vivo

Supplemental_Data.zip

The compressed Supplemental_Data.zip archive contains two files: 1. Supplemental Data_S3, the complete HIP HOP profiles are provided as tab delimited .txt files. The sensitivity and z-scores for all HIP strains present in the pool across all tested compounds are listed in the HIP-exp-scores-annotation.txt file, scores for all HOP strains present in the pool are listed in the HOP-exp-scores-annotation.txt file. To reproduce the plots depicted in the article the sensitivity can be plotted on the y-axis, the z-score on the x-axis. Each gene is annotated with systematic name, common name, description, GO category etc. 2. The Supplemental_Data_S4.zip file containing the in silico docking solution. It is provided as a standard Protein Data Bank file named 1N0H_compound1_docked.pdb. UNK1 refers to the docked compound 1, all other structures are labeled as in the original 1N0H.pdb file downloaded from RCSB protein databank www.pdb.org

Identification and Evaluation of Novel Acetolactate Synthase Inhibitors as Antifungal Agents

High-throughput phenotypic screening against the yeast Saccharomyces cerevisiae revealed a series of triazolopyrimidine-sulfonamide compounds with broad-spectrum antifungal activity, no significant cytotoxicity, and low protein binding. To elucidate the target of this series, we have applied a chemogenomic profiling approach using the S. cerevisiae deletion collection. All compounds of the series yielded highly similar profiles that suggested acetolactate synthase (Ilv2p, which catalyzes the first common step in branched-chain amino acid biosynthesis) as a possible target. The high correlation with profiles of known Ilv2p inhibitors like chlorimuron-ethyl provided further evidence for a similar mechanism of action. Genome-wide mutagenesis in S. cerevisiae identified 13 resistant clones with 3 different mutations in the catalytic subunit of acetolactate synthase that also conferred cross-resistance to established Ilv2p inhibitors. Mapping of the mutations into the published Ilv2p crystal structure outlined the chlorimuron-ethyl binding cavity, and it was possible to dock the triazolopyrimidine-sulfonamide compound into this pocket in silico. However, fungal growth inhibition could be bypassed through supplementation with exogenous branched-chain amino acids or by the addition of serum to the medium in all of the fungal organisms tested except for Aspergillus fumigatus. Thus, these data support the identification of the triazolopyrimidine-sulfonamide compounds as inhibitors of acetolactate synthase but suggest that targeting may be compromised due to the possibility of nutrient bypass in vivo