UDP-sugar substrates of HAS3 regulate its O-GlcNAcylation, intracellular traffic, extracellular shedding and correlate with melanoma progression

Cell Mol Life Sci. 2016 Aug;73(16):3183-204. doi: 10.1007/s00018-016-2158-5. Epub 2016 Feb 16.

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.

Keywords: 4MU; GNPDA; Glucosamine; Mannose; OGT; UGDH.

MeSH terms

  • Acetylglucosamine / metabolism
  • Acylation
  • Animals
  • COS Cells
  • Cell Line
  • Cell Line, Tumor
  • Chlorocebus aethiops
  • Disease Progression
  • Endocytosis
  • Glucuronosyltransferase / metabolism*
  • Humans
  • Hyaluronan Synthases
  • Hyaluronic Acid / metabolism*
  • Melanoma / metabolism*
  • Melanoma / pathology
  • Protein Transport
  • Skin / metabolism*
  • Skin / pathology
  • Skin Neoplasms / metabolism*
  • Skin Neoplasms / pathology
  • Uridine Diphosphate N-Acetylglucosamine / metabolism
  • Uridine Diphosphate Sugars / metabolism*

Substances

  • Uridine Diphosphate Sugars
  • Uridine Diphosphate N-Acetylglucosamine
  • Hyaluronic Acid
  • Glucuronosyltransferase
  • HAS1 protein, human
  • HAS3 protein, human
  • Hyaluronan Synthases
  • Acetylglucosamine