Phosphatidylinositol 3-kinases Play a Suppressive Role in Electrotaxis of Dictyostelium Cells Through Akt and ERK
10.16476/j.pibb.2023.0084
- VernacularTitle:磷脂酰肌醇3激酶通过Akt和ERK抑制盘基网柄菌细胞的趋电性
- Author:
Xiao-Xue GE
1
;
Rui-Da JIANG
1
;
Xiao-Yan WANG
1
;
Jing GAO
2
;
Run-Chi GAO
1
Author Information
1. School of Life Science, Yunnan Normal University, Kunming 650500, China
2. School of Agriculture, Yunnan University, Kunming 650500, China
- Publication Type:Journal Article
- Keywords:
cell migration;
CRISPR/Cas9;
electrotaxis;
Dictyostelium discoideum
- From:
Progress in Biochemistry and Biophysics
2024;51(3):685-695
- CountryChina
- Language:Chinese
-
Abstract:
ObjectivePhosphatidylinositol 3 kinases (PI3Ks) play an important role in cell directional movement by regulating F-actin. However, the structure and function of PI3Ks are complex. The role of PI3Ks in cell electrotaxis is not fully understood. Therefore, in this study, the model organism Dictyostelium discoideum cells were used as experimental materials to explore the role of PI3K1 and PI3K2 in electrotaxis. MethodsFirstly, PI3K1 coding gene pikA knockout mutant and PI3K2 coding gene pikB knockout mutant were constructed by CRISPR/Cas9 system. Secondly, two mutants were placed in a DC electric field with a strength of 12 V/cm and the electrotaxis were analyzed. ResultsData analysis showed that the direction index of wild-type cells in DC electric field was (0.86±0.03), while the direction index of pikA- and pikB- mutants in DC electric field was (0.95±0.02) and (0.94±0.03), respectively. In addition, the average trajectory speed of wild-type cells in the electric field was (3.34±0.08) μm/min, while the average trajectory speed of pikA- and pikB- mutants were (4.85±0.20) μm/min and (5.48±0.15) μm/min, respectively. The t test showed that there were significant differences in the directedness index and speed between the mutant and wild type. Western blot results showed that both phosphorylated Akt and phosphorylated ERK were significantly increased in pikA- and pikB- mutants. ConclusionPI3K1 and PI3K2 may inhibit the electrotaxis of Dictyostelium discoideum cells by increasing the activity of Akt and ERK.
