Different Location of Katanin P60 Effect on Cellular Microtubule Cutting
Volume 7, Issue 1, June 2019, Pages: 10-13
Received: Aug. 12, 2019;
Accepted: Aug. 28, 2019;
Published: Sep. 19, 2019
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Li Shaojin, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Luo Jianxian, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Ji Zhisheng, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Yang Yuhao, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Zhang Guowei, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Tan Minghui, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
Yang Hua, Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China
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Katanin is a heterodimeric hydrolase belonging to the AAA protein family and its function is cutting microtubules. It plays an important role in cell division, neural development, cell migration, and the formation of motor organelles. Katanin has two subunits including katanin p80 and p60. This research is aimed to study the effect of different location of katanin p60 on cellular microtubule cutting. First, katanin p60 gene was amplified by PCR, and the eukaryotic recombinant plasmid GFP-katanin p60 and GFP- katanin p60M1S were constructed. The recombinant plasmid was identified by enzyme digestion and sequencing. The expression of katanin in RFL-6 cells was detected by western blot analysis. And, the distribution of katanin p60 in RFL-6 cells and its effect on microtubule were further examined. The results showed that the GFP-katanin p60 and GFP-katanin p60M1S recombinant plasmids was successfully constructed. By western blot analysis, GFP-katanin p60 and GFP-katanin p60M1S recombinant plasmids could be successfully expressed in RFL-6 cells. The microtubule cutting ability of GFP-katanin p60M1S which distribute in whole-cell was stronger than GFP-katanin p60 which distribute in nuclear. Therefore, the cutting function on microtubule of katanin p60 depends on its distribution, and the whole cell distribution is beneficial to katanin p60 in cutting microtubule.
Katanin p60, Microtubule Cutting, RFL-6 Cell, Protein Distribution
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Different Location of Katanin P60 Effect on Cellular Microtubule Cutting, Cell Biology.
Vol. 7, No. 1,
2019, pp. 10-13.
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