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Cargo binding and regulatory sites in the tail of fungal conventional kinesin

Nature Cell Biology volume 2, pages 333–338 (2000)Cite this article

Abstract

Here, using a quantitative in vivo assay, we map three regions in the carboxy terminus of conventional kinesin that are involved in cargo association, folding and regulation, respectively. Using C-terminal and internal deletions, point mutations, localization studies, and an engineered ‘minimal’ kinesin, we identify five heptads of a coiled-coil domain in the kinesin tail that are necessary and sufficient for cargo association. Mutational analysis and in vitro ATPase assays highlight a conserved motif in the globular tail that is involved in regulation of the motor domain; a region preceding this motif participates in folding. Although these sites are spatially and functionally distinct, they probably cooperate during activation of the motor for cargo transport.

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Figure 1: Rescue of the kinesin-null phenotype and immunolocalization of Myc-tagged kinesin constructs.
Figure 2: Point mutations in the cargo-binding region, and ‘minimal’ kinesin.
Figure 3: Mutants exhibiting kinesin accumulation at hyphal tips.
Figure 4: Summary of the functional features of kinesin tail domains and the effects of the mutations studied here.

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Acknowledgements

We thank S. Fuchs for technical assistance and U. Euteneuer for critical reading of the manuscript. This work was supported by the Deutsche Forschungsgemeinschaft, the Volkswagen Stiftung and the Fonds der Chemischen Industrie.

Correspondence and requests for materials should be addressed to M.S.

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  1. Adolf Butenandt Institute, Cell Biology, University of Munich, Munich, 80336, Germany

    Stephan Seiler, Jochen Kirchner, Christian Horn, Athina Kallipolitou, Günther Woehlke & Manfred Schliwa

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Correspondence to Manfred Schliwa.

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Seiler, S., Kirchner, J., Horn, C. et al. Cargo binding and regulatory sites in the tail of fungal conventional kinesin. Nat Cell Biol 2, 333–338 (2000). https://doi.org/10.1038/35014022

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