biostudies-literatureKhandelwal NKU.S. Department of Health & Human Services | NIH | National Institute of General Medical SciencesNIAID NIH HHSNIH HHSDivision of Intramural Research, National Institute of Allergy and Infectious DiseasesNIGMS NIH HHS1278https://www.ebi.ac.uk/biostudies/studies/S-EPMC8917219biostudies-literatureUnknown13(1)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 strucNature communicationsThe structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation.PMC8917219R01AI156270R00GM11424U24 GM129547R00 GM114245R01 AI156270S10 OD011981Millan CRZangari SITomasiak TMAvila SWilliams DThaker TMKhandelwal NKfalseThe structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation.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 struc2022-01-01T00:00:00Z2022 Mar2026-05-10T01:38:23.699Z2025-02-19T01:11:56.178ZS-EPMC89172193527748710.1038/s41467-022-28811-w