Danhelovska 2018 MiP2018
Acyl-CoA binding domain containing 3 (ACBD3) is a protein of Golgi apparatus (GA) which has many functions in the cell such as division of neuronal cells, in neurodegeneration, in lipid homeostasis, in cell stress response, in apoptosis or in maintenance of GA. Moreover, ACBD3 has important role in replication of some plus RNA viruses. In steroidogenic cells ACBD3 is essential in transport of cholesterol into mitochondria. In this type of cells, ACBD3 coexists in multiprotein complex named transduceosome together with other proteins (VDAC1, TSPO, PKA regulatory subunits α (PKAR1α) etc.) in the outer mitochondrial membrane. ACBD3 works as a scaffold protein of TSPO and PKAR1α. Knockdown of ACBD3 suppresses hormone-induced steroidogenesis .
In patient with homozygous mutation in ACBD3 gene, we observed different distribution of cholesterol in fibroblasts and increased and enlarged mitochondria in muscle together with decreased amount of OXPHOS complexes CI, CIII, CIV and ATP synthase.
The aim of study is to characterize the impact of full absence of ACBD3 in HeLa and HEK293 cells. We create ACBD3 knockout (KO) cell lines by CRISPR/CAS9 system to study overall impact of the protein absence on mitochondrial metabolism, using protein and functional analyzes of the OXPHOS system or microscopic analyzes.
From our preliminary results in KO HeLa cells, altered cholesterol distribution and decreased OXPHOS capacity measured by Oroboros (Oxygraph-2k) were found. Reduced overall respiration was also detected by Seahorse Bioanalyzer.
Labels: MiParea: Respiration, Genetic knockout;overexpression
Organism: Human Tissue;cell: HEK, HeLa
Coupling state: OXPHOS
- Dept Pediatrics Adolescent Medicine, First Fac Medicine, Charles Univ General Univ Hospital, Prague, Czech Republic. - [email protected]
References and Support
- Li H, Degenhardt B, Tobin D, Yao ZX, Tasken K, Papadopoulos V (2001) Identification, localization, and function in steroidogenesis of PAP7: a peripheral-type benzodiazepine receptor- and PKA (RIalpha)-associated protein. Mol Endocrinol 15:2211–28.
Supported by research projects: GAUK 542217, GAČR 14-36804G, AZV 16-31932A, RVO VFN64165/2012 and SVV 260367.