Flexible microtubule anchoring modulates the bi-directional motility of the kinesin-5 Cin8

© 2021. The Author(s), under exclusive licence to Springer Nature Switzerland AG..

Two modes of motility have been reported for bi-directional kinesin-5 motors: (a) context-dependent directionality reversal, a mode in which motors undergo persistent minus-end directed motility at the single-molecule level and switch to plus-end directed motility in different assays or under different conditions, such as during MT gliding or antiparallel sliding or as a function of motor clustering; and (b) bi-directional motility, defined as movement in two directions in the same assay, without persistent unidirectional motility. Here, we examine how modulation of motor-microtubule (MT) interactions affects these two modes of motility for the bi-directional kinesin-5, Cin8. We report that the large insert in loop 8 (L8) within the motor domain of Cin8 increases the MT affinity of Cin8 in vivo and in vitro and is required for Cin8 intracellular functions. We consistently found that recombinant purified L8 directly binds MTs and L8 induces single Cin8 motors to behave according to context-dependent directionality reversal and bi-directional motility modes at intermediate ionic strength and according to a bi-directional motility mode in an MT surface-gliding assay under low motor density conditions. We propose that the largely unstructured L8 facilitates flexible anchoring of Cin8 to the MTs. This flexible anchoring enables the direct observation of bi-directional motility in motility assays. Remarkably, although L8-deleted Cin8 variants exhibit a strong minus-end directed bias at the single-molecule level, they also exhibit plus-end directed motility in an MT-gliding assay. Thus, L8-induced flexible MT anchoring is required for bi-directional motility of single Cin8 molecules but is not necessary for context-dependent directionality reversal of Cin8 in an MT-gliding assay.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:78

Enthalten in:

Cellular and molecular life sciences : CMLS - 78(2021), 16 vom: 10. Aug., Seite 6051-6068

Sprache:

Englisch

Beteiligte Personen:

Pandey, Himanshu [VerfasserIn]
Singh, Sudhir Kumar [VerfasserIn]
Sadan, Mayan [VerfasserIn]
Popov, Mary [VerfasserIn]
Singh, Meenakshi [VerfasserIn]
Davidov, Geula [VerfasserIn]
Inagaki, Sayaka [VerfasserIn]
Al-Bassam, Jawdat [VerfasserIn]
Zarivach, Raz [VerfasserIn]
Rosenfeld, Steven S [VerfasserIn]
Gheber, Larisa [VerfasserIn]

Links:

Volltext

Themen:

Bi-directional motility
CIN8 protein, S cerevisiae
Cin8
Context-dependent directionality reversal
EC 3.6.4.4
Journal Article
Kinesin-5
Kinesins
Motor–microtubule interaction
Saccharomyces cerevisiae Proteins

Anmerkungen:

Date Completed 02.08.2021

Date Revised 04.12.2021

published: Print-Electronic

Citation Status MEDLINE

doi:

10.1007/s00018-021-03891-x

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM328203491