The Acyl-CoA-binding domain-containing protein (ACBD3) plays multiple roles across the cell. Although generally associated with the Golgi apparatus, it operates also in mitochondria. In steroidogenic cells, ACBD3 is an important part of a multiprotein complex transporting cholesterol into mitochondria. Balance in mitochondrial cholesterol is essential for proper mitochondrial protein biosynthesis, among others. We generated ACBD3 knock-out (ACBD3-KO) HEK293 and HeLa cells and characterized the impact of protein absence on mitochondria, Golgi, and lipid profile. In ACBD3-KO cells, cholesterol level and mitochondrial structure and functions are not altered, demonstrating that an alternative pathway of cholesterol transport into mitochondria exists. However, ACBD3-KO cells exhibit enlarged Golgi area with absence of stacks and ribbon-like formation, confirming the importance of ACBD3 in Golgi stacking. The glycosylation of the LAMP2 glycoprotein was not affected by the altered Golgi structure. Moreover, decreased sphingomyelins together with normal ceramides and sphingomyelin synthase activity reveal the importance of ACBD3 in ceramide transport from ER to Golgi.

• Keywords
• ACBD3, Golgi, OXPHOS, cholesterol, knock-out, mitochondria,
• MeSH
• Adaptor Proteins, Signal Transducing metabolism   MeSH
• Biological Transport physiology   MeSH
• Ceramides metabolism   MeSH
• Cholesterol metabolism   MeSH
• Glycosylation MeSH
• Golgi Apparatus metabolism   MeSH
• HEK293 Cells MeSH
• HeLa Cells MeSH
• Humans MeSH
• Membrane Proteins metabolism   MeSH
• Lysosomal-Associated Membrane Protein 2 metabolism   MeSH
• Mitochondria metabolism   MeSH
• Signal Transduction physiology   MeSH
• Transferases (Other Substituted Phosphate Groups) metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• ACBD3 protein, human MeSH Browser
• Adaptor Proteins, Signal Transducing MeSH
• Ceramides MeSH
• Cholesterol MeSH
• Membrane Proteins MeSH
• Lysosomal-Associated Membrane Protein 2 MeSH
• phosphatidylcholine-ceramide phosphocholine transferase MeSH Browser
• Transferases (Other Substituted Phosphate Groups) MeSH