Nucleotide-dependent interaction of the N-terminal domain of MukB with microtubules.
The MukB protein from Escherichia coli has a domain structure that is reminiscent of the eukaryotic motor proteins kinesin and myosin: N-terminal globular domains, a region of coiled-coil, and a specialised C-terminal domain. Sequence alignment of the N-terminal domain of MukB with the kinesin motor domain indicated an approximately 22% sequence identity. These observations raised the possibility that MukB might be a prokaryotic motor protein and, due to the sequence homology shared with kinesin, might bind to microtubules (Mts). We found that a construct encoding the first 342 residues of MukB (Muk342) binds specifically to Mts and shares a number of properties with the motor domain of kinesin. Visualisation of the Muk342 decorated Mt complexes using negative stain electron microscopy indicated that the Muk342 smoothly decorates the outside of Mts. Biochemical data demonstrate that Muk342 decorates Mts with a binding stoichiometry of one Muk342 monomer per tubulin monomer. These findings strongly suggest that MukB has a role in force generation and that it is a prokaryotic homologue of kinesin and myosin.
Lockhart
A
A
Structural Studies Division, MRC-Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, United Kingdom. andrew.lockhart@pharma.Novartis.com
Kendrick-Jones
J
J
D000227:Adenine Nucleotides / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000648:ultrastructure; D001665:Binding Sites; D002868:Chromosomal Proteins, Non-Histone; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins; D004330:Drosophila; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D006801:Humans; D016547:Kinesins / Q000737:chemistry; D008870:Microtubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000227:Adenine Nucleotides; D001426:Bacterial Proteins; D002868:Chromosomal Proteins, Non-Histone; D003598:Cytoskeletal Proteins; D029968:Escherichia coli Proteins; C071525:FtsZ protein, Bacteria; C067035:MukB protein, E coli; D011994:Recombinant Proteins; D016547:Kinesins