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Biophys J. 2015 Dec 15;109(12):2546-2561. doi: 10.1016/j.bpj.2015.11.011.

Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails.

Author information

1
Molecular and Cellular Biology Graduate Program, University of Massachusetts-Amherst, Amherst, Massachusetts; Department of Physics, University of Massachusetts-Amherst, Amherst, Massachusetts.
2
Program in Physical Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
3
Department of Physics, University of Massachusetts-Amherst, Amherst, Massachusetts. Electronic address: rossj@physics.umass.edu.

Abstract

Microtubule dynamics in cells are regulated by associated proteins that can be either stabilizers or destabilizers. A class of destabilizers that is important in a large number of cellular activities is the microtubule-severing enzymes, yet little is known about how they function. Katanin p60 was the first ATPase associated with microtubule severing. Here, we investigate the activity of katanin severing using a GFP-labeled human version. We quantify the effect of katanin concentration on katanin binding and severing activity. We find that free tubulin can inhibit severing activity by interfering with katanin binding to microtubules. The inhibition is mediated by the sequence of the tubulin and specifically depends on the carboxy-terminal tails. We directly investigate the inhibition effect of tubulin carboxy-terminal tails using peptide sequences of α-, β-, or detyrosinated α-tubulin tails that have been covalently linked to bovine serum albumin. Our results show that β-tubulin tails are the most effective at inhibiting severing, and that detyrosinated α-tubulin tails are the least effective. These results are distinct from those for other severing enzymes and suggest a scheme for regulation of katanin activity in cells dependent on free tubulin concentration and the modification state of the tubulin.

PMID:
26682813
PMCID:
PMC4699919
DOI:
10.1016/j.bpj.2015.11.011
[Indexed for MEDLINE]
Free PMC Article

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