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Understanding the internalization dynamics of N‐acetylglucosamine transporter (Ngt1) in Candida albicans
Author(s) -
Rao Kongara Hanumantha,
Ghosh Swagata
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.654.2
Subject(s) - internalization , endocytosis , microbiology and biotechnology , biology , transporter , candida albicans , ubiquitin , lipid raft , cell membrane , cytosol , cell , chemistry , biochemistry , signal transduction , gene , enzyme
Integral membrane proteins are subjected to tight regulation in response to environmental cues. Ubiquitylation mediated endocytosis forms an integral part of this machinery. However, such a dynamics has not been worked out for the novel N‐acetylglucosamine transporter (Ngt1) in the pathogenic yeast Candida albicans . N‐acetylglucosamine (GlcNAc) stimulates a wide range of signalling events in several organisms apart from evoking the phenomenon of virulence in several pathogens. Therefore, immense importance lies in the proper understanding of its uptake before the initiation of signalling. In this study we have investigated the Ngt1 PostTranslationalModifications (PTMs) and its interaction with membrane lipids during internalization on cell membrane. We employed western blot, FACS, lipid binding assay, LC‐MS/MS and mutant analysis to understand the various key events involved in internalization. Our studies demonstrate that there exists sophisticated and subtle phosphorylation dependent ubiquitylation machinery that is responsible for proper maintenance of the transporter level at the cell surface. Further our study highlights how the transporter interacts with components of membrane microdomains (lipid rafts) like ergosterol for modulation of the transporter activity. Being a robust transporter, it can provide a tool for development of an inducible promoter and can help in designing therapeutic reagents to modulate GlcNAc entry. Support or Funding Information DST‐SERB (Early Career Research Award)., Government of India., India Localization dynamics of GlcNAc transporter (Ngt1) upon glucose additionAt 90 min of GlcNAc induction almost all the fraction of Ngt1‐GFP resides in the cell membrane. Immediately within 5 min after addition of glucose some fraction protein which internalized into cytosol appeared as dotted punctate like structures (Early‐endosomes, late‐endosomes, MultiVesicularBodies, TransGolgi) and within 30–60 min, the degraded fraction appeared in vacuole, merged with vacuolar dye, CMAC CellTracker Blue.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .