The structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation
Year: 2022
Type: article
Abstract: Yeast Cadmium Factor 1 (Ycf1) sequesters heavy metals and glutathione into the vacuole to counter cell stress. Ycf1 belongs to the ATP binding cassette C-subfamily (ABCC) of transporters, many of which are regulated by phosphorylation on intrinsically-disordered domains. The regulatory mechanism of phosphorylation is still poorly understood. Here, we report two cryo-EM structures of Ycf1 at 3.4 Å and 4.0 Å resolution in inward-facing open conformations that capture previously unobserved ordered states of the intrinsically disordered regulatory domain (R-domain). R-domain phosphorylation is clearly evident and induces a topology promoting electrostatic and hydrophobic interactions with Nucleotide Binding Domain 1 (NBD1) and the Lasso motif. These interactions stay constant between the structures and are related by rigid body movements of the NBD1/R-domain complex. Biochemical data further show R-domain phosphorylation reorganizes the Ycf1 architecture and is required for maximal ATPase activity. Together, we provide insights into how R-domains control ABCC transporter activity.
Source: Nature Communications
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $5,890
Funders: National Science Foundation, U.S. Department of Energy, U.S. Department of Health and Human Services, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, Oregon Health and Science University, Arizona State University, University of Arizona, National Institutes of Health, Office of Science, Eyring Materials Center, Arizona State University, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, Biological and Environmental Research
Awards: Structural mechanisms for ABC multidrug transporter selectivity and transport ABC, G1083383770, G1213529597, G2691348929, G3508161151, G4429615956, G5520316217, G5718375046, Pacific Northwest Center for Cryo-EM, G6780141363, G7562546725, G7673727097, G7811164366, G7913850483, JEOL JEM-1400 Transmission Electron Microscope, ATP Binding Cassette (ABC) Transporters in Fungal Drug Tolerance
OpenAlex ID: W4221071430