15 Å Resolution Model of the Monomeric Kinesin Motor, KIF1A

A two-headed structure has been widely believed to be essential for the kinesin molecular motor to move processively on the track, microtubules. However, we have recently demonstrated that a monomeric motor domain construct of KIF1A (C351), a kinesin superfamily protein, moves processively, taking about 700 steps before being detached from microtubules. To elucidate the mechanism of its single-headed processivity, we examined the C351 -MT interaction by mutant analysis and high-resolution cryo-EM. Mutant analysis indicated the importance of a highly positively charged loop, the "K loop," for such processivity. A 15 A resolution structure unambiguously docked with the available atomic models revealed "K loop" as an extra microtubule-binding domain specific to KIF1A, and bound to the C terminus of tubulin. The site-specific cross-linking further confirmed this model.

[1]  E. Mandelkow,et al.  Image Reconstructions of Microtubules Decorated with Monomeric and Dimeric Kinesins: Comparison with X-Ray Structure and Implications for Motility , 1998, The Journal of cell biology.

[2]  D. Chrétien,et al.  Relationship between moiré patterns, tubulin shape, and microtubule polarity. , 1998, Cell motility and the cytoskeleton.

[3]  N. Hirokawa,et al.  The neuron-specific kinesin superfamily protein KIF1A is a uniqye monomeric motor for anterograde axonal transport of synaptic vesicle precursors , 1995, Cell.

[4]  E. Nogales,et al.  High-Resolution Model of the Microtubule , 1999, Cell.

[5]  E. Egelman,et al.  An algorithm for straightening images of curved filamentous structures. , 1986, Ultramicroscopy.

[6]  D. DeRosier,et al.  Reconstruction of three-dimensional images from electron micrographs of structures with helical symmetry. , 1970, Journal of molecular biology.

[7]  N. Hirokawa,et al.  Defect in Synaptic Vesicle Precursor Transport and Neuronal Cell Death in KIF1A Motor Protein–deficient Mice , 1998, The Journal of cell biology.

[8]  A. Hoenger,et al.  Three different approaches for calculating the three-dimensional structure of microtubules decorated with kinesin motor domains. , 1997, Journal of structural biology.

[9]  Ronald D. Vale,et al.  Crystal structure of the kinesin motor domain reveals a structural similarity to myosin , 1996, Nature.

[10]  E A Merritt,et al.  Raster3D Version 2.0. A program for photorealistic molecular graphics. , 1994, Acta crystallographica. Section D, Biological crystallography.

[11]  K Schulten,et al.  Nucleotide-dependent movements of the kinesin motor domain predicted by simulated annealing. , 1998, Biophysical journal.

[12]  R. Beroukhim,et al.  Distortion correction of tubular crystals: improvements in the acetylcholine receptor structure. , 1997, Ultramicroscopy.

[13]  N. Hirokawa,et al.  Three-dimensional structure of the kinesin head-microtubule complex. , 1995, Nature.

[14]  N. Hirokawa,et al.  Direct visualization of the microtubule lattice seam both in vitro and in vivo , 1994, The Journal of cell biology.

[15]  Ronald D. Vale,et al.  Direction determination in the minus-end-directed kinesin motor ncd , 1998, Nature.

[16]  E. Mandelkow,et al.  The Crystal Structure of Dimeric Kinesin and Implications for Microtubule-Dependent Motility , 1997, Cell.

[17]  Andreas Hoenger,et al.  A Model for the Microtubule-Ncd Motor Protein Complex Obtained by Cryo-Electron Microscopy and Image Analysis , 1997, Cell.

[18]  S. Fuller,et al.  Determination of microtubule polarity by cryo-electron microscopy. , 1996, Structure.

[19]  K. Hirose,et al.  Three-dimensional cryoelectron microscopy of dimeric kinesin and ncd motor domains on microtubules. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[20]  D. Chrétien,et al.  New data on the microtubule surface lattice , 1991, Biology of the cell.

[21]  R. Wade,et al.  Nucleotide-dependent conformations of the kinesin dimer interacting with microtubules. , 1998, Structure.

[22]  M. Whittaker,et al.  Molecular structure of F-actin and location of surface binding sites , 1990, Nature.

[23]  K. Hirose,et al.  Congruent docking of dimeric kinesin and ncd into three-dimensional electron cryomicroscopy maps of microtubule-motor ADP complexes. , 1999, Molecular biology of the cell.

[24]  Ronald D Vale,et al.  The Directional Preference of Kinesin Motors Is Specified by an Element outside of the Motor Catalytic Domain , 1997, Cell.

[25]  Marin van Heel,et al.  Similarity measures between images , 1987 .

[26]  N. Hirokawa,et al.  Three-dimensional structure of the kinesin headá¤-microtubule complex , 1995, Nature.

[27]  Kenneth H. Downing,et al.  Structure of the αβ tubulin dimer by electron crystallography , 1998, Nature.

[28]  Y. Berwald‐Netter,et al.  Developmental regulation of polyglutamylated alpha- and beta-tubulin in mouse brain neurons. , 1994, Journal of cell science.

[29]  N. Unwin,et al.  Ion channel of acetylcholine receptor reconstructed from images of postsynaptic membranes , 1988, Nature.

[30]  J. Mccammon,et al.  Situs: A package for docking crystal structures into low-resolution maps from electron microscopy. , 1999, Journal of structural biology.

[31]  T Yasunaga,et al.  Extensible and object-oriented system Eos supplies a new environment for image analysis of electron micrographs of macromolecules. , 1996, Journal of structural biology.

[32]  N. Hirokawa,et al.  Kinesin and dynein superfamily proteins in organelle transport and cell division. , 1998, Current opinion in cell biology.

[33]  R. Wade,et al.  A model of the microtubule–kinesin complex based on electron cryomicroscopy and X-ray crystallography , 1998, Current Biology.

[34]  K. Hirose,et al.  Nucleotide-dependent angular change in kinesin motor domain bound to tubulin , 1995, Nature.

[35]  E. Mandelkow,et al.  X-ray structure of motor and neck domains from rat brain kinesin. , 1997, Biochemistry.

[36]  N. Hirokawa,et al.  KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria , 1994, Cell.

[37]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[38]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[39]  N. Hirokawa,et al.  A processive single-headed motor: kinesin superfamily protein KIF1A. , 1999, Science.

[40]  N. Hirokawa,et al.  Kinesin and dynein superfamily proteins and the mechanism of organelle transport. , 1998, Science.