Acetyl-CoA carboxylase 1 is the rate-limiting enzyme of fatty acid synthesis. It carboxylates acetyl-CoA to form malonyl-CoA in an ATP-dependent manner. It was reported that acetyl-CoA carboxylase 1 was concentrated in the center of the region occupied by chromosomes and partly overlapped with chromosomes and spindle fibers in mitotic prophase. After knocking down acetyl-CoA carboxylase 1 in HCT116 cells, the rate of chromosome segregation defects increased significantly. The aim of this study was to investigate the molecular function of acetyl-CoA carboxylase 1 in terms of its enzymatic activity, polymerization ability, mitotic prophase localization, and role in regulating chromosome segregation during mitosis. To track acetyl-CoA carboxylase 1 in living cells, a monomeric enhanced green fluorescent protein tag was fused to the N-terminus of human acetyl-CoA carboxylase 1 isoform 4. According to the results from affinity purification, enzymatic assay, immuno-fluorescence, and live-cell imaging, it was hypothesized that the green fluorescent protein tagging abolished the catalytic activity of acetyl-CoA carboxylase 1 and made the enzyme bind exclusively to the nuclear envelope.
|Date of Award||Jul 2023|
|Original language||English (US)|
- Biological, Environmental Sciences and Engineering
|Supervisor||Wolfgang Fischle (Supervisor)|