Feedback Microtubule Control and Microtubule-Actin Cross-talk in Revealed by Integrative Proteomic and Cell Biology Analysis of Mutants.

Mol Cell Proteomics

From the ‡Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů 27, 783 71 Olomouc, Czech Republic;

Published: September 2017

Microtubule organization and dynamics are critical for key developmental processes such as cell division, elongation, and morphogenesis. Microtubule severing is an essential regulator of microtubules and is exclusively executed by KATANIN 1 in In this study, we comparatively studied the proteome-wide effects in two mutants. Thus, shotgun proteomic analysis of roots and aerial parts of single nucleotide mutant and T-DNA insertion mutant was carried out. We have detected 42 proteins differentially abundant in both and KATANIN 1 dysfunction altered the abundance of proteins involved in development, metabolism, and stress responses. The differential regulation of tubulins and microtubule-destabilizing protein MDP25 implied a feedback microtubule control in mutants. Furthermore, deregulation of profilin 1, actin-depolymerizing factor 3, and actin 7 was observed. These findings were confirmed by immunoblotting analysis of actin and by microscopic observation of actin filaments using fluorescently labeled phalloidin. Results obtained by quantitative RT-PCR analysis revealed that changed protein abundances were not a consequence of altered expression levels of corresponding genes in the mutants. In conclusion, we show that abundances of several cytoskeletal proteins as well as organization of microtubules and the actin cytoskeleton are amended in accordance with defective microtubule severing.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5587860PMC
http://dx.doi.org/10.1074/mcp.M117.068015DOI Listing

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