Study of the Effect of N-terminal mEGFP Tagging on the Enzymatic Activity and Cellular Localization of Human Acetyl-CoA Carboxylase 1

  • Linsheng Zhang

Student thesis: Master's Thesis


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 AwardJul 2023
Original languageEnglish (US)
Awarding Institution
  • Biological, Environmental Sciences and Engineering
SupervisorWolfgang Fischle (Supervisor)

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