Abstract:
In
Schizosaccharomyces pombe cells, the mitochondrial dynamain-related protein Dnm1 is a key protein that regulates the dynamic processes of mitochondrial division and fusion. To study the effects of
dnm1 gene deletion on mitosis and energy metabolism in fission yeast cells, live cell imaging technology was used to analyze the changes in cell mitosis kinetics, RT-qPCR technology was used to analyze the transcription levels of
cdc2 and
cdc13 genes, high performance liquid chromatography-mass spectrometry technology was used to detect the changes in energy metabolites and validated them. The results indicate that the deletion of
dnm1 gene could lead to slow growth of fission yeast cells, and live cell imaging shows that the length of microtubule bundles in the
dnm1Δ strain increased by (0.83±0.70) μm (**
P<0.01) compared to the wild-type during the interphase of mitosis, the number of strains producing 5 microtubule bundles increased, while the number of strains producing 3 microtubule bundles decreased. The spindle elongation time in the
dnm1Δ strain was prolonged by (0.85±0.02) minutes (*
P<0.05) in the mitosis prophase, and was prolonged by (5.8±1.62) minutes (**
P<0.01) in the mitosis anaphase. The spindle elongation speed slowed down by (0.06±0.00) μm/min (*
P<0.05) in the mitosis anaphase. The
dnm1Δ strain also exhibited delayed spindle breakage and delayed chromosome segregation. The results of high performance liquid chromatography-mass spectrometry technology indicated that the
dnm1Δ strain had a deficiency in coenzyme synthesis, resulting in a significant decrease in NADPH content (*
P<0.05). The relative content of intermediate metabolites such as glucose 6-phosphate, fructose 6-phosphate, citric acid, cis aconitic acid, pyruvate, isocitrate, and L-malic acid were significantly reduced (*
P<0.05), there was also a defect of ATP production. The verification results showed that the metabolite analysis results were reliable and and the expression level of
cdc2 gene in the logarithmic growth stage of the
dnm1Δ strain was significantly lower than that of the wild-type. This study provided a certain scientific basis for further exploring the function and related molecular mechanisms of Dnm1 protein in cell mitosis.