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

• Published in: Cellular and molecular life sciences : CMLS 2016-08, Vol.73 (16), p.3183-3204
• Main Authors: Deen, Ashik Jawahar, Arasu, Uma Thanigai, Pasonen-Seppänen, Sanna, Hassinen, Antti, Takabe, Piia, Wojciechowski, Sara, Kärnä, Riikka, Rilla, Kirsi, Kellokumpu, Sakari, Tammi, Raija, Tammi, Markku, Oikari, Sanna
• Format: Article
• Language: English
• Subjects: Acetylglucosamine - metabolism; Acylation; Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Cancer; Cell Biology; Cell Line; Cell Line, Tumor; Chlorocebus aethiops; COS Cells; Disease Progression; Endocytosis; Enzymes; Glucuronosyltransferase - metabolism; Humans; Hyaluronan Synthases; Hyaluronic Acid - metabolism; Life Sciences; Melanoma; Melanoma - metabolism; Melanoma - pathology; Membranes; Molecular biology; Original; Original Article; Plasma; Protein Transport; Skin - metabolism; Skin - pathology; Skin Neoplasms - metabolism; Skin Neoplasms - pathology; Sugar; Uridine Diphosphate N-Acetylglucosamine - metabolism; Uridine Diphosphate Sugars - metabolism
• ISSN: 1420-682X; 1420-9071
• DOI: 10.1007/s00018-016-2158-5

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.