title: UDP-sugar substrates of HAS3 regulate its O-GlcNAcylation, intracellular traffic, extracellular shedding and correlate with melanoma progression.
abstract: Hyaluronan content is a powerful prognostic factor in many cancer types, but the molecular basis of its synthesis in cancer still remains unclear. Hyaluronan synthesis requires the transport of hyaluronan synthases (HAS1-3) from Golgi to plasma membrane (PM), where the enzymes are activated. For the very first time, the present study demonstrated a rapid recycling of HAS3 between PM and endosomes, controlled by the cytosolic levels of the HAS substrates UDP-GlcUA and UDP-GlcNAc. Depletion of UDP-GlcNAc or UDP-GlcUA shifted the balance towards HAS3 endocytosis, and inhibition of hyaluronan synthesis. In contrast, UDP-GlcNAc surplus suppressed endocytosis and lysosomal decay of HAS3, favoring its retention in PM, stimulating hyaluronan synthesis, and HAS3 shedding in extracellular vesicles. The concentration of UDP-GlcNAc also controlled the level of O-GlcNAc modification of HAS3. Increasing O-GlcNAcylation reproduced the effects of UDP-GlcNAc surplus on HAS3 trafficking, while its suppression showed the opposite effects, indicating that O-GlcNAc signaling is associated to UDP-GlcNAc supply. Importantly, a similar correlation existed between the expression of GFAT1 (the rate limiting enzyme in UDP-GlcNAc synthesis) and hyaluronan content in early and deep human melanomas, suggesting the association of UDP-sugar metabolism in initiation of melanomagenesis. In general, changes in glucose metabolism, realized through UDP-sugar contents and O-GlcNAc signaling, are important in HAS3 trafficking, hyaluronan synthesis, and correlates with melanoma progression.
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland. ashik.jawahardeen@uef.fi.
authors.forename: Ashik Jawahar
authors.initials: AJ
authors.lastname: Deen
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Uma Thanigai
authors.initials: UT
authors.lastname: Arasu
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Sanna
authors.initials: S
authors.lastname: Pasonen-Seppänen
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90014, Oulu, Finland.
authors.forename: Antti
authors.initials: A
authors.lastname: Hassinen
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Piia
authors.initials: P
authors.lastname: Takabe
authors.affiliation: A. I. Virtanen Institute for Molecular Sciences, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Sara
authors.initials: S
authors.lastname: Wojciechowski
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Riikka
authors.initials: R
authors.lastname: Kärnä
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Kirsi
authors.initials: K
authors.lastname: Rilla
authors.affiliation: Faculty of Biochemistry and Molecular Medicine, University of Oulu, 90014, Oulu, Finland.
authors.forename: Sakari
authors.initials: S
authors.lastname: Kellokumpu
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Raija
authors.initials: R
authors.lastname: Tammi
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland.
authors.forename: Markku
authors.initials: M
authors.lastname: Tammi
authors.affiliation: Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland. sanna.oikari@uef.fi.|Institute of Dentistry, School of Medicine, University of Eastern Finland, 70210, Kuopio, Finland. sanna.oikari@uef.fi.
authors.forename: Sanna
authors.initials: S
authors.lastname: Oikari
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000215:Acylation; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D018450:Disease Progression; D004705:Endocytosis; D014453:Glucuronosyltransferase / Q000378:metabolism*; D006801:Humans; D000076002:Hyaluronan Synthases; D006820:Hyaluronic Acid / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D021381:Protein Transport; D012867:Skin / Q000378:metabolism* / Q000473:pathology; D012878:Skin Neoplasms / Q000378:metabolism* / Q000473:pathology; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism; D014539:Uridine Diphosphate Sugars / Q000378:metabolism*
chemical_list: D014539:Uridine Diphosphate Sugars; D014537:Uridine Diphosphate N-Acetylglucosamine; D006820:Hyaluronic Acid; D014453:Glucuronosyltransferase; C523794:HAS1 protein, human; C105458:HAS3 protein, human; D000076002:Hyaluronan Synthases; D000117:Acetylglucosamine
keywords: 4MU; GNPDA; Glucosamine; Mannose; OGT; UGDH