PT  - JOURNAL ARTICLE
AU  - Graham M Burkart
AU  - Ram Dixit
TI  - Microtubule bundling by MAP65-1 protects against severing by inhibiting the binding of katanin
AID  - 10.1101/520445
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 520445
4099  - http://biorxiv.org/content/early/2019/01/14/520445.short
4100  - http://biorxiv.org/content/early/2019/01/14/520445.full
AB  - The microtubule-severing enzyme katanin regulates the organization and turnover of microtubule arrays by the localized breakdown of microtubule polymers. In land plants, katanin (KTN1) activity is essential for the formation of linearly organized cortical microtubule arrays which determine the axis of cell expansion. Cell biological studies have shown that even though KTN1 binds to the sidewalls of single and bundled microtubules, severing activity is restricted to microtubule crossover and nucleation sites, indicating that cells contain protective mechanisms to prevent indiscriminate microtubule severing. Here, we show that the microtubule bundling protein MAP65-1 inhibits KTN1-mediated microtubule severing in vitro. Severing is inhibited at bundled microtubule segments and the severing rate of non-bundled microtubules is reduced by MAP65-1 in a concentration-dependent manner. Using various MAP65-1 mutant proteins, we demonstrate that efficient crosslinking of microtubules is crucial for this protective effect and that microtubule binding alone is not sufficient. Reduced severing due to microtubule bundling by MAP65-1 correlated to decreased binding of KTN1 to these microtubules. Taken together, our work reveals that crosslinking of microtubules by MAP65-1 confers resistance to severing by inhibiting the binding of katanin and identifies the structural features of MAP65-1 that are important for this activity.Highlight Summary Cortical microtubule bundles resist severing in vivo. Here, we show that crosslinking of microtubules by MAP65-1 inhibits severing in a dose-dependent manner by preventing katanin from binding to these microtubules.AbbreviationsAse1p,anaphase spindle elongation protein 1;GFP,green fluorescent protein;KTN1,katanin;MAP2,microtubule-associated protein 2;MAP65,microtubule-associated protein of 65 kDa size;PRC1,protein regulator of cytokinesis 1.