Structural analysis of the ZEN-4/CeMKLP1 motor domain and its interaction with microtubules.
The centralspindlin complex is required for the assembly and maintenance of the central spindle during late anaphase and the completion of cytokinesis. It is composed of two copies each of the kinesin-like protein ZEN-4, a Caenorhabditis elegans MKLP-1 (Kinesin-6 family), and the RhoGAP CYK-4. By using cryo-electron microscopy and helical 3D reconstruction, we are investigating the structural features of the interactions between monomeric and dimeric motor domain constructs of ZEN-4 and microtubules. We have calculated helically averaged 3D maps of microtubules decorated with ZEN-4 motor domain in the presence of AMP-PNP, ADP, ADP-AlF(4)(-), and nucleotide-free conditions. We used statistical difference mapping to compare these maps among each other and to related maps obtained from microtubules decorated with a well-characterized Kinesin-1 motor domain from Neurospora crassa. Thereby, we found distinct structural features in microtubule-ZEN-4 complexes that may directly relate to the functional properties of ZEN-4 and centralspindlin. Furthermore, we investigated the location, structure, and function of a highly conserved extension of approximately 50 residues unique to the Kinesin-6 subfamily, located in the motor core loop6/beta4 region.
Hizlan
Dilem
D
European Molecular Biology Laboratory, Structural Biology and Biocomputing Programme, Heidelberg, Germany.
Mishima
Masanori
M
Tittmann
Peter
P
Gross
Heinz
H
Glotzer
Michael
M
Hoenger
Andreas
A
D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000378:metabolism; D017124:Conserved Sequence; D020285:Cryoelectron Microscopy; D019281:Dimerization; D005656:Fungal Proteins / Q000737:chemistry; D021621:Imaging, Three-Dimensional; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism; D008432:Mathematical Computing; D008870:Microtubules / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000737:chemistry; D009942:Organometallic Compounds / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D040681:Structural Homology, Protein
D029742:Caenorhabditis elegans Proteins; D005656:Fungal Proteins; D009942:Organometallic Compounds; C114455:ZEN-4 protein, C elegans; C105164:adenosine diphosphate tetrafluoroaluminate; D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D016547:Kinesins