Abstract
Knowledge on the distribution and dynamics of glycosylation enzymes in the Golgi is essential for better understanding this modification. Here, using a combination of CRISPR/Cas9 knockin technology and super-resolution microscopy, we show that the Golgi complex is assembled by a number of small 'Golgi units' that have 1-3 μm in diameter. Each Golgi unit contains small domains of glycosylation enzymes which we call 'zones'. The zones of N- and O-glycosylation enzymes are colocalised. However, they are less colocalised with the zones of a glycosaminoglycan synthesizing enzyme. Golgi units change shapes dynamically and the zones of glycosylation enzymes rapidly move near the rim of the unit. Photobleaching analysis indicates that a glycosaminoglycan synthesizing enzyme moves between units. Depletion of giantin dissociates units and prevents the movement of glycosaminoglycan synthesizing enzymes, which leads to insufficient glycosaminoglycan synthesis. Thus, we show the structure-function relationship of the Golgi and its implications in human pathogenesis.
© 2024. The Author(s).
MeSH terms
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CRISPR-Cas Systems
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Glycosaminoglycans* / metabolism
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Glycosylation
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Golgi Apparatus* / metabolism
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Golgi Matrix Proteins
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HeLa Cells
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Humans
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
Substances
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Glycosaminoglycans
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macrogolgin
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Membrane Proteins
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Golgi Matrix Proteins
Grants and funding
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17H0622/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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21H02658/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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21K06734/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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17H06413/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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17H06420/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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18H05275/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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18H05275/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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17H06413/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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17H06420/Ministry of Education, Culture, Sports, Science and Technology (MEXT)
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18H05275/Ministry of Education, Culture, Sports, Science and Technology (MEXT)