Volume 7, Issue 1, June 2019, Page: 10-13
Different Location of Katanin P60 Effect on Cellular Microtubule Cutting
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
Received: Aug. 12, 2019;       Accepted: Aug. 28, 2019;       Published: Sep. 19, 2019
DOI: 10.11648/j.cb.20190701.12      View  30      Downloads  12
Abstract
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.
Keywords
Katanin p60, Microtubule Cutting, RFL-6 Cell, Protein Distribution
To cite this article
Li Shaojin, Luo Jianxian, Ji Zhisheng, Yang Yuhao, Zhang Guowei, Tan Minghui, Yang Hua, Different Location of Katanin P60 Effect on Cellular Microtubule Cutting, Cell Biology. Vol. 7, No. 1, 2019, pp. 10-13. doi: 10.11648/j.cb.20190701.12
Copyright
Copyright © 2019 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Frickey T, Lupas AN. Phylogenetic analysis of AAA proteins. Journal of structural biology. 2004; 146 (1-2): 2-10.
[2]
Hartman JJ, Mahr J, McNally K, et al. Katanin, a microtubule-severing protein, is a novel AAA ATPase that targets to the centrosome using a WD40-containing subunit. Cell. 1998; 93 (2): 277-287.
[3]
McNally FJ, Vale RD. Identification of katanin, an ATPase that severs and disassembles stable microtubules. Cell. 1993; 75 (3): 419-429.
[4]
McNally KP, Bazirgan OA, McNally FJ. Two domains of p80 katanin regulate microtubule severing and spindle pole targeting by p60 katanin. Journal of cell science. 2000; 113 (Pt 9): 1623-1633.
[5]
Ghosh DK, Dasgupta D, Guha A. Models, Regulations, and Functions of Microtubule Severing by Katanin. ISRN molecular biology. 2012; 2012: 596289.
[6]
Davis LJ, Odde DJ, Block SM, et al. The importance of lattice defects in katanin-mediated microtubule severing in vitro. Biophysical journal. 2002; 82 (6): 2916-2927.
[7]
Mohrbach H, Kulic IM. Motor driven microtubule shape fluctuations: force from within the lattice. Physical review letters. 2007; 99 (21): 218102.
[8]
Allard JF, Wasteneys GO, Cytrynbaum EN. Mechanisms of self-organization of cortical microtubules in plants revealed by computational simulations. Molecular biology of the cell. 2010; 21 (2): 278-286.
[9]
Baas PW, Karabay A, Qiang L. Microtubules cut and run. Trends in cell biology. 2005; 15 (10): 518-524.
[10]
Sudo H, Baas PW. Acetylation of microtubules influences their sensitivity to severing by katanin in neurons and fibroblasts. The Journal of neuroscience : the official journal of the Society for Neuroscience. 2010; 30 (21): 7215-7226.
[11]
Fletcher DA, Mullins RD. Cell mechanics and the cytoskeleton. Nature. 2010; 463 (7280): 485-492.
[12]
Zhang D, Rogers GC, Buster DW, et al. Three microtubule severing enzymes contribute to the "Pacman-flux" machinery that moves chromosomes. The Journal of cell biology. 2007; 177 (2): 231-242.
[13]
Bailey ME, Sackett DL, Ross JL. Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails. Biophysical journal. 2015; 109 (12): 2546-2561.
[14]
Tao J, Feng C, Rolls MM. The microtubule-severing protein fidgetin acts after dendrite injury to promote their degeneration. 2016; 129 (17): 3274-3281.
[15]
Taylor JL, White SR, Lauring B, et al. Crystal structure of the human spastin AAA domain. Journal of structural biology. 2012; 179 (2): 133-137.
[16]
Austin TO, Matamoros AJ, Friedman JM, et al. Nanoparticle Delivery of Fidgetin siRNA as a Microtubule-based Therapy to Augment Nerve Regeneration. Scientific reports. 2017; 7 (1): 9675.
[17]
Karabay A, Yu W, Solowska JM, et al. Axonal growth is sensitive to the levels of katanin, a protein that severs microtubules. The Journal of neuroscience : the official journal of the Society for Neuroscience. 2004; 24 (25): 5778-5788.
Browse journals by subject