ER-Golgi dynamics of HS-modifying enzymes via vesicular trafficking is a critical prerequisite for the delineation of HS biosynthesis

• Published in: Carbohydrate polymers 2021-03, Vol.255, p.117477, Article 117477
• Main Authors: Meneghetti, Maria C.Z., Deboni, Paula, Palomino, Carlos M.V., Braga, Luiz P., Cavalheiro, Renan P., Viana, Gustavo M., Yates, Edwin A., Nader, Helena B., Lima, Marcelo A.
• Format: Article
• Language: English
• Subjects: Biological Transport - drug effects; Biosynthesis; Brefeldin A - pharmacology; Cell Membrane - chemistry; Cell Membrane - drug effects; Cell Membrane - enzymology; COP-Coated Vesicles - enzymology; COP-Coated Vesicles - genetics; COPI; COPII; Endoplasmic Reticulum - chemistry; Endoplasmic Reticulum - drug effects; Endoplasmic Reticulum - enzymology; Gene Expression Regulation; Golgi apparatus; Golgi Apparatus - chemistry; Golgi Apparatus - drug effects; Golgi Apparatus - enzymology; Heparan sulfate; Heparin - pharmacology; Heparitin Sulfate - biosynthesis; Human Umbilical Vein Endothelial Cells - cytology; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - enzymology; Humans; Plasmids - chemistry; Plasmids - metabolism; Primary Cell Culture; Transfection; Vesicular Transport Proteins - genetics; Vesicular Transport Proteins - metabolism
• ISSN: 0144-8617; 1879-1344
• DOI: 10.1016/j.carbpol.2020.117477

•HS-modifying enzymes are actively engaged in both anterograde and retrograde Golgi transport.•HS-modifying enzymes are sorted and trafficked by similar mechanisms.•BFA treatment changed HS-modifying enzymes vesicular trafficking.•HS-modifying enzymes moved from cis to trans-Golgi during heparin stimulus.•Changes in the localization of HS-modifying enzymes correlate with changes in HS structure. The cell surface and extracellular matrix polysaccharide, heparan sulfate (HS) conveys chemical information to control crucial biological processes. HS chains are synthesized in a non-template driven process mainly in the Golgi apparatus, involving a large number of enzymes capable of subtly modifying its substitution pattern, hence, its interactions and biological effects. Changes in the localization of HS-modifying enzymes throughout the Golgi were found to correlate with changes in the structure of HS, rather than protein expression levels. Following BFA treatment, the HS-modifying enzymes localized preferentially in COPII vesicles and at the trans-Golgi. Shortly after heparin treatment, the HS-modifying enzyme moved from cis to trans-Golgi, which coincided with increased HS sulfation. Finally, it was shown that COPI subunits and Sec24 gene expression changed. Collectively, these findings demonstrate that knowledge of the ER-Golgi dynamics of HS-modifying enzymes via vesicular trafficking is a critical prerequisite for the complete delineation of HS biosynthesis.