A structural analysis of the interaction between ncd tail and tubulin protofilaments.
ncd is a minus-end directed, kinesin-like motor, which binds to microtubules with its motor domain and its cargo domain as well. Typical of retrograde motors, the motor domain of ncd locates to the C-terminal end of the polypeptide chain, and hence, the cargo domain constitutes the N-terminal region. To date, several studies have investigated the interaction properties of the motor domain with microtubules, but very few structural data are available about the tail itself or its interaction with microtubules as cargo. Here, we applied cryo-electron microscopy and helical 3D image reconstruction to 15 protofilament microtubules decorated with an ncd tail fragment (N-terminal residues 83-187, named NT6). In our study, the ncd tail shows a behaviour resembling filamentous MAPs such as tau protein, exhibiting a highly flexible structure with no large globular domains. NT6 binds to four different sites on the outer side of microtubules within the proximity of the kinesin motor-binding site. Two of these sites locate within the groove between two neighbouring protofilaments, and appear as strong binding sites, while the other two sites, located at the outer rim, appear to play a secondary role. In addition, the ncd tail fragment induces the formation of large protofilament sheets, suggesting a tail-induced modification of lateral protofilament contacts.
Wendt
Thomas
T
European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Karabay
Arzu
A
Krebs
Angelika
A
Gross
Heinz
H
Walker
Richard
R
Hoenger
Andreas
A
D020285:Cryoelectron Microscopy; D029721:Drosophila Proteins; D021621:Imaging, Three-Dimensional; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure
D029721:Drosophila Proteins; D014404:Tubulin; C066696:ncd protein, Drosophila; D016547:Kinesins