GRASP55 regulates intra-Golgi localization of glycosylation enzymes to control glycosphingolipid biosynthesis

EMBO J. 2021 Oct 18;40(20):e107766. doi: 10.15252/embj.2021107766. Epub 2021 Sep 13.

Abstract

The Golgi apparatus, the main glycosylation station of the cell, consists of a stack of discontinuous cisternae. Glycosylation enzymes are usually concentrated in one or two specific cisternae along the cis-trans axis of the organelle. How such compartmentalized localization of enzymes is achieved and how it contributes to glycosylation are not clear. Here, we show that the Golgi matrix protein GRASP55 directs the compartmentalized localization of key enzymes involved in glycosphingolipid (GSL) biosynthesis. GRASP55 binds to these enzymes and prevents their entry into COPI-based retrograde transport vesicles, thus concentrating them in the trans-Golgi. In genome-edited cells lacking GRASP55, or in cells expressing mutant enzymes without GRASP55 binding sites, these enzymes relocate to the cis-Golgi, which affects glycosphingolipid biosynthesis by changing flux across metabolic branch points. These findings reveal a mechanism by which a matrix protein regulates polarized localization of glycosylation enzymes in the Golgi and controls competition in glycan biosynthesis.

Keywords: GRASP55; Golgi apparatus; glucosylceramide synthase; glycosphingolipids; glycosylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism
  • Autoantigens / genetics
  • Autoantigens / metabolism
  • Brefeldin A / pharmacology
  • Ceramides / metabolism
  • Cholera Toxin / pharmacology
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism
  • Gene Expression
  • Glycosphingolipids / metabolism*
  • Glycosylation / drug effects
  • Golgi Apparatus / drug effects
  • Golgi Apparatus / genetics
  • Golgi Apparatus / metabolism*
  • Golgi Matrix Proteins / genetics
  • Golgi Matrix Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism
  • Shiga Toxin / pharmacology

Substances

  • Adaptor Proteins, Signal Transducing
  • Autoantigens
  • Ceramides
  • Cytoskeletal Proteins
  • GORASP1 protein, human
  • GORASP2 protein, human
  • Glycosphingolipids
  • Golgi Matrix Proteins
  • Golgin subfamily A member 2
  • Membrane Proteins
  • PLEKHA8 protein, human
  • TRIP11 protein, mouse
  • macrogolgin
  • Brefeldin A
  • Shiga Toxin
  • Cholera Toxin
  • CERT1 protein, human
  • Protein Serine-Threonine Kinases