Cytosolic N- and C-Termini of the Aspergillus nidulans FurE Transporter Contain Distinct Elements that Regulate by Long-Range Effects Function and Specificity. Issue 19 (6th September 2019)

• Record Type: Journal Article
• Authors: Papadaki, Georgia F.; Lambrinidis, George; Zamanos, Andreas; Mikros, Emmanuel; Diallinas, George
• Is Part Of: Journal of molecular biology. Volume 431:Issue 19(2019)
• Page Start: 3827
• Page End: 3844
• Publication Date: 2019-09-06
• Subjects: Aspergillus nidulans -- fungi -- transport -- folding -- membrane lipids; NCS1 nucleobase cation symporter 1 -- TMS transmembrane α-helical segment -- MD molecular dynamics -- LID loop interacting domain -- 5FU 5-fluorouracil; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.07.013
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: FurE, a member of the NCS1 family, is an Aspergillus nidulans transporter specific for uracil, allantoin and uric acid. Recently, we showed that C- or N-terminally truncated FurE versions are blocked for endocytosis and surprisingly show modified substrate specificities. Bifluorescence complementation assays and genetic analyses supported the idea that C- and N-termini interact dynamically and through this interaction regulate selective substrate translocation. Here we functionally dissect and define distinct motifs crucial for endocytosis, transport activity, substrate specificity and folding, in both cytosolic termini of FurE. Subsequently, we obtain novel genetic and in silico evidence indicating that the molecular dynamics of specific N- and C-terminal regions exert long-range effects on the gating mechanism responsible for substrate selection, via pH-dependent interactions with other internal cytosolic loops and membrane lipids. Our work shows that expanded cytoplasmic termini, acquired through evolution mostly in eukaryotic transporters, provide novel specific functional roles. Graphical abstract: Unlabelled Image