"Gene Ontology annotation through association of InterPro records with GO terms"

"Annotation inferences using phylogenetic trees"

"Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping"

"Here we provide evidence that yeast RFT1 encodes an evolutionarily conserved protein required for the translocation of Man5GlcNAc2-PP-Dol from the cytoplasmic to the lumenal leaflet of the ER membrane."

"A young patient diagnosed with a congenital disorder of glycosylation characterized by an intracellular accumulation of DolPP-GlcNAc(2)Man(5) was found to carry a homozygous point mutation in the RFT1 gene."

"Despite the low sequence similarity between the yeast and the human RFT1 proteins, we demonstrated both their functional orthology and the pathologic effect of the human p.R67C mutation by complementation assay in Deltarft1 yeast cells."

"The causality of the RFT1 p.R67C mutation was further established by restoration of normal glycosylation profiles in patient-derived fibroblasts after lentiviral expression of a normal RFT1 cDNA."

"RFT1 deficiency in both yeast and human cells leads to the accumulation of incomplete DolPP-GlcNAc(2)Man(5) and to a profound glycosylation disorder in humans."

"We now show that a specific ER protein(s), but not Rft1, is required to flip Man(5)GlcNAc(2)-PP-Dol in reconstituted vesicles."

"Rft1 may have a critical accessory role in translocating Man(5)GlcNAc(2)-PP-Dol in vivo, but the Man(5)GlcNAc(2)-PP-Dol flippase itself remains to be identified."

"Specific transbilayer translocation of dolichol-linked oligosaccharides by an endoplasmic reticulum flippase"

"no difference was seen in the level of M5-DLO flippase activity in sealed wild type and Rft1-depleted microsomal vesicles"

"although Rft1 may play a critical role in vivo, depletion of this protein does not impair the transbilayer movement of M5-DLO in sealed microsomal fractions prepared from disrupted cells."

"The pathogenic character of the novel mutations was illustrated by the accumulation of Man(5)GlcNAc(2)-PP-dolichol and by reduced recombinant DNase 1 secretion."

"The clinical phenotype of these patients comprised typical CDG symptoms in addition to sensorineural deafness, rarely reported in CDG patients."

"We report that TbRft1-null procyclic trypanosomes grow nearly normally."

"They have normal steady-state levels of mDLO and significant N-glycosylation, indicating robust M5-DLO flippase activity."

"rather than facilitating M5-DLO flipping, Rft1 facilitates conversion of M5-DLO to mDLO by another mechanism, possibly by acting as an M5-DLO chaperone."

"Here we present a human 'all-by-all' reference interactome map of human binary protein interactions, or 'HuRI'."

"As with ScRft1, proteoliposomes reconstituted with HsRft1 showed almost 90% conversion of M5GN2 to M3GN2 after 2 h of incubation."

"Translocation was ATP-independent, occurred with similar kinetics for both dolichol or phytanol lipid carriers, and importantly, was dependent on addition of purified Rft1 or archaeal Agl23"

"we sought to reconstitute liposomes with purified ScRft1 or HhAgl23 in reactions that contained α1-2 mannosidase, without ATP"

"Genetic studies identified Rft1 as the M5GN2-PP-Dol flippase in vivo but are at odds with biochemical data suggesting Rft1 is dispensable for flipping in vitro."

"We show that it is a multispanning membrane protein located in the ER, with its N and C termini facing the cytoplasm."

"It is therefore not known what essential role Rft1 plays in N-glycosylation."

"other studies indicated that Rft1 is not required for heptasaccharide lipid flipping in microsomes or unilamellar vesicles reconstituted with ER membrane proteins, nor is it required for the viability of at least one eukaryote."

"While this activity has been demonstrated in liposomes reconstituted with purified Rft1, biochemical evidence of additional M5-DLO scramblases and the viability of Rft1-null Trypanosoma brucei suggest that scrambling may be a moonlighting function of Rft1 rather than its essential cellular role."

"Strikingly, the portal-blocking mutant which is predicted to lack scramblase activity supported robust growth."

"These data suggest that while M5-DLO binding is important for Rft1's essential function, scrambling activity is dispensable."

"We speculate that Rft1's essential role may be as an M5-DLO chaperone, capturing and routing M5-DLO propitiously on the cytoplasmic side of the ER to coordinate DLO biosynthesis."

"central-cavity-lining transmembrane (TM) positions are the single most evolutionarily constrained structural class, well above the TM background, whereas lateral lipid-portal-lining positions are statistically indistinguishable from background"

"they are 28% charged and 47% polar, the chemistry of substrate coordination"

"the 15 most-constrained TM residues have a mean pairwise distance of 19.2 Å vs a random-TM null of 30.1 Å"

"PF04506/RFT1 has no alignable archaeal/bacterial members"

"my first (ray-tracing) portal detector found"

"E298 points inward, not outward"

"R67 is inward-facing but peripheral"

"a polytopic ~12–14-TM, two-lobed helical bundle enclosing a discrete central pore axis that is geometrically separable from the lipid-facing periphery"

"is consistent with the MOP/MATE/MurJ two-domain architecture independently modelled for Rft1"

"an invariant Arg across all orthologs"

"the apo human model's innermost lining is net-acidic"

"No disease residue lines the lateral dolichol portal."

"model, so cavity-localised disease mutations do not discriminate them."

"Human RFT1 localizes throughout the ER. DeepTMHMM and experimental tagging/topology mapping indicate ~14 transmembrane spans with both N- and C-termini facing the cytosol"