Distinct Interaction Modes of the Kinesin-13 Motor Domain with the Microtubule

Chandrima Chatterjee, Matthieu P M H Benoit, Vania DePaoli, Juan D. Diaz-Valencia, Ana B. Asenjo, Gary J. Gerfen, David J. Sharp, Hernando J. Sosa

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Kinesins-13s are members of the kinesin superfamily of motor proteins that depolymerize microtubules (MTs) and have no motile activity. Instead of generating unidirectional movement over the MT lattice, like most other kinesins, kinesins-13s undergo one-dimensional diffusion (ODD) and induce depolymerization at the MT ends. To understand the mechanism of ODD and the origin of the distinct kinesin-13 functionality, we used ensemble and single-molecule fluorescence polarization microscopy to analyze the behavior and conformation of Drosophila melanogaster kinesin-13 KLP10A protein constructs bound to the MT lattice. We found that KLP10A interacts with the MT in two coexisting modes: one in which the motor domain binds with a specific orientation to the MT lattice and another where the motor domain is very mobile and able to undergo ODD. By comparing the orientation and dynamic behavior of mutated and deletion constructs we conclude that 1) the Kinesin-13 class specific neck domain and loop-2 help orienting the motor domain relative to the MT. 2) During ODD the KLP10A motor-domain changes orientation rapidly (rocks or tumbles). 3) The motor domain alone is capable of undergoing ODD. 4) A second tubulin binding site in the KLP10A motor domain is not critical for ODD. 5) The neck domain is not the element preventing KLP10A from binding to the MT lattice like motile kinesins.

Original languageEnglish (US)
Pages (from-to)1593-1604
Number of pages12
JournalBiophysical Journal
Volume110
Issue number7
DOIs
StatePublished - Apr 12 2016

Fingerprint

Kinesin
Microtubules
Neck
Polarization Microscopy
Microtubule Proteins
Fluorescence Polarization
Tubulin
Drosophila melanogaster
Fluorescence Microscopy
Binding Sites

ASJC Scopus subject areas

  • Biophysics

Cite this

Distinct Interaction Modes of the Kinesin-13 Motor Domain with the Microtubule. / Chatterjee, Chandrima; Benoit, Matthieu P M H; DePaoli, Vania; Diaz-Valencia, Juan D.; Asenjo, Ana B.; Gerfen, Gary J.; Sharp, David J.; Sosa, Hernando J.

In: Biophysical Journal, Vol. 110, No. 7, 12.04.2016, p. 1593-1604.

Research output: Contribution to journalArticle

Chatterjee, Chandrima ; Benoit, Matthieu P M H ; DePaoli, Vania ; Diaz-Valencia, Juan D. ; Asenjo, Ana B. ; Gerfen, Gary J. ; Sharp, David J. ; Sosa, Hernando J. / Distinct Interaction Modes of the Kinesin-13 Motor Domain with the Microtubule. In: Biophysical Journal. 2016 ; Vol. 110, No. 7. pp. 1593-1604.
@article{4160378076314995b54b0eea1c0518eb,
title = "Distinct Interaction Modes of the Kinesin-13 Motor Domain with the Microtubule",
abstract = "Kinesins-13s are members of the kinesin superfamily of motor proteins that depolymerize microtubules (MTs) and have no motile activity. Instead of generating unidirectional movement over the MT lattice, like most other kinesins, kinesins-13s undergo one-dimensional diffusion (ODD) and induce depolymerization at the MT ends. To understand the mechanism of ODD and the origin of the distinct kinesin-13 functionality, we used ensemble and single-molecule fluorescence polarization microscopy to analyze the behavior and conformation of Drosophila melanogaster kinesin-13 KLP10A protein constructs bound to the MT lattice. We found that KLP10A interacts with the MT in two coexisting modes: one in which the motor domain binds with a specific orientation to the MT lattice and another where the motor domain is very mobile and able to undergo ODD. By comparing the orientation and dynamic behavior of mutated and deletion constructs we conclude that 1) the Kinesin-13 class specific neck domain and loop-2 help orienting the motor domain relative to the MT. 2) During ODD the KLP10A motor-domain changes orientation rapidly (rocks or tumbles). 3) The motor domain alone is capable of undergoing ODD. 4) A second tubulin binding site in the KLP10A motor domain is not critical for ODD. 5) The neck domain is not the element preventing KLP10A from binding to the MT lattice like motile kinesins.",
author = "Chandrima Chatterjee and Benoit, {Matthieu P M H} and Vania DePaoli and Diaz-Valencia, {Juan D.} and Asenjo, {Ana B.} and Gerfen, {Gary J.} and Sharp, {David J.} and Sosa, {Hernando J.}",
year = "2016",
month = "4",
day = "12",
doi = "10.1016/j.bpj.2016.02.029",
language = "English (US)",
volume = "110",
pages = "1593--1604",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "7",

}

TY - JOUR

T1 - Distinct Interaction Modes of the Kinesin-13 Motor Domain with the Microtubule

AU - Chatterjee, Chandrima

AU - Benoit, Matthieu P M H

AU - DePaoli, Vania

AU - Diaz-Valencia, Juan D.

AU - Asenjo, Ana B.

AU - Gerfen, Gary J.

AU - Sharp, David J.

AU - Sosa, Hernando J.

PY - 2016/4/12

Y1 - 2016/4/12

N2 - Kinesins-13s are members of the kinesin superfamily of motor proteins that depolymerize microtubules (MTs) and have no motile activity. Instead of generating unidirectional movement over the MT lattice, like most other kinesins, kinesins-13s undergo one-dimensional diffusion (ODD) and induce depolymerization at the MT ends. To understand the mechanism of ODD and the origin of the distinct kinesin-13 functionality, we used ensemble and single-molecule fluorescence polarization microscopy to analyze the behavior and conformation of Drosophila melanogaster kinesin-13 KLP10A protein constructs bound to the MT lattice. We found that KLP10A interacts with the MT in two coexisting modes: one in which the motor domain binds with a specific orientation to the MT lattice and another where the motor domain is very mobile and able to undergo ODD. By comparing the orientation and dynamic behavior of mutated and deletion constructs we conclude that 1) the Kinesin-13 class specific neck domain and loop-2 help orienting the motor domain relative to the MT. 2) During ODD the KLP10A motor-domain changes orientation rapidly (rocks or tumbles). 3) The motor domain alone is capable of undergoing ODD. 4) A second tubulin binding site in the KLP10A motor domain is not critical for ODD. 5) The neck domain is not the element preventing KLP10A from binding to the MT lattice like motile kinesins.

AB - Kinesins-13s are members of the kinesin superfamily of motor proteins that depolymerize microtubules (MTs) and have no motile activity. Instead of generating unidirectional movement over the MT lattice, like most other kinesins, kinesins-13s undergo one-dimensional diffusion (ODD) and induce depolymerization at the MT ends. To understand the mechanism of ODD and the origin of the distinct kinesin-13 functionality, we used ensemble and single-molecule fluorescence polarization microscopy to analyze the behavior and conformation of Drosophila melanogaster kinesin-13 KLP10A protein constructs bound to the MT lattice. We found that KLP10A interacts with the MT in two coexisting modes: one in which the motor domain binds with a specific orientation to the MT lattice and another where the motor domain is very mobile and able to undergo ODD. By comparing the orientation and dynamic behavior of mutated and deletion constructs we conclude that 1) the Kinesin-13 class specific neck domain and loop-2 help orienting the motor domain relative to the MT. 2) During ODD the KLP10A motor-domain changes orientation rapidly (rocks or tumbles). 3) The motor domain alone is capable of undergoing ODD. 4) A second tubulin binding site in the KLP10A motor domain is not critical for ODD. 5) The neck domain is not the element preventing KLP10A from binding to the MT lattice like motile kinesins.

UR - http://www.scopus.com/inward/record.url?scp=84963610653&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84963610653&partnerID=8YFLogxK

U2 - 10.1016/j.bpj.2016.02.029

DO - 10.1016/j.bpj.2016.02.029

M3 - Article

VL - 110

SP - 1593

EP - 1604

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 7

ER -