Cryo-EM of dynein microtubule-binding domains shows how an axonemal dynein distorts the microtubule

Abstract
Data availability
Article and author information
Metrics

Abstract

Dyneins are motor proteins responsible for transport in the cytoplasm and the beating of axonemes in cilia and flagella. They bind and release microtubules via a compact microtubule-binding domain (MTBD) at the end of a coiled-coil stalk. We address how cytoplasmic and axonemal dynein MTBDs bind microtubules at near atomic resolution. We decorated microtubules with MTBDs of cytoplasmic dynein-1 and axonemal dynein DNAH7 and determined their cryo-EM structures using helical Relion. The majority of the MTBD is rigid upon binding, with the transition to the high affinity state controlled by the movement of a single helix at the MTBD interface. DNAH7 contains an 18-residue insertion, found in many axonemal dyneins, that contacts the adjacent protofilament. Unexpectedly we observe that DNAH7, but not dynein-1, induces large distortions in the microtubule cross-sectional curvature. This raises the possibility that dynein coordination in axonemes is mediated via conformational changes in the microtubule.

Data availability

CryoEM maps have been deposited to the EMDB and are instructed to be released on publication under codes 10060 and 10061PDB models have been deposited to the PDB and are instructed to be released on publication under codes 6RZA and 6RZB

The following data sets were generated

1. Lacey SE
2. He S
3. Scheres SHW
4. Carter AP
(2019) CryoEM maps
EMDataBank, EMD-10060.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-10060
1. Lacey SE
2. He S
3. Scheres SHW
4. Carter AP
(2019) CryoEM maps
EMDataBank, EMD-10061.

http://www.ebi.ac.uk/pdbe/entry/emdb/EMD-10061
1. Lacey SE
2. He S
3. Scheres SHW
4. Carter AP
(2019) PDB models
Protein Data Bank, 6RZA.

http://www.rcsb.org/structure/6RZA
1. Lacey SE
2. He S
3. Scheres SHW
4. Carter AP
(2019) PDB models
Protein Data Bank, 6RZB.

http://www.rcsb.org/structure/6RZB

The following previously published data sets were used

1. Zhang R
2. Nogales E
(2015) EMPIAR dataset of EB3 decorated MTs
EMPIAR, 10030.

https://www.ebi.ac.uk/pdbe/emdb/empiar/entry/10030/

Article and author information

Author details

Samuel E Lacey

MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.
Shaoda He

MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.
Sjors HW Scheres

MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
Sjors HW Scheres, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0002-0462-6540
Andrew P Carter

MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

For correspondence
cartera@mrc-lmb.cam.ac.uk

Competing interests
Andrew P Carter, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0001-7292-5430

Funding

Wellcome (WT210711)

Andrew P Carter

Medical Research Council (MC_UP_A025_1011)

Andrew P Carter

Medical Research Council (MC_UP_A025_1011)

Sjors HW Scheres

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.