Journal Article

Structural basis of mechano-chemical coupling by the mitotic kinesin KIF14

Asenjo, Ana B., Dhakal, Sabin, Paydar, Mohammadjavad, Benoit, Matthieu P.M.H., Kwok, Benjamin H., Sosa, Hernando

2021

Overview

KIF14 is a mitotic kinesin whose malfunction is associated with cerebral and renal developmental defects and several cancers. Like other kinesins, KIF14 couples ATP hydrolysis and microtubule binding to the generation of mechanical work, but the coupling mechanism between these processes is still not fully clear. Here we report 20 high-resolution (2.7–3.9 Å) cryo-electron microscopy KIF14-microtubule structures with complementary functional assays. Analysis procedures were implemented to separate coexisting conformations of microtubule-bound monomeric and dimeric KIF14 constructs. The data provide a comprehensive view of the microtubule and nucleotide induced KIF14 conformational changes. It shows that: 1) microtubule binding, the nucleotide species, and the neck-linker domain govern the transition between three major conformations of the motor domain; 2) an undocked neck-linker prevents the nucleotide-binding pocket to fully close and dampens ATP hydrolysis; 3) 13 neck-linker residues are required to assume a stable docked conformation; 4) the neck-linker position controls the hydrolysis rather than the nucleotide binding step; 5) the two motor domains of KIF14 dimers adopt distinct conformations when bound to the microtubule; and 6) the formation of the two-heads-bound-state introduces structural changes in both motor domains of KIF14 dimers. These observations provide the structural basis for a coordinated chemo-mechanical kinesin translocation model. KIF14 is a mitotic kinesin whose malfunction is associated with cerebral and renal developmental defects and several cancers. Here the authors use cryoEM to determine 20 structures of KIF14 constructs bound to microtubules in the presence of different nucleotide analogues and provide the structural basis for a coordinated chemo-mechanical kinesin translocation model.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 101/28 / 631/535/1258/1259 / 631/80/128/1923 / Adenosine Diphosphate - metabolism / Adenosine triphosphate / Adenosine Triphosphate - metabolism / Animals / Binding / Binding Sites / Cancer / Conformation / Coupling / Cryoelectron Microscopy / Datasets / Defects / Dimers / Domains / Electron microscopy / Humanities and Social Sciences / Hydrolysis / Kinesin / Kinesins - chemistry / Kinesins - genetics / Kinesins - metabolism / Ligands / Mechanical properties / Mice / Microscopy / Microtubules / Microtubules - chemistry / Microtubules - genetics / Microtubules - metabolism / Molecular Docking Simulation / multidisciplinary / Neck / Nucleotide analogs / Nucleotides / Oncogene Proteins - chemistry / Oncogene Proteins - genetics / Oncogene Proteins - metabolism / Protein Binding / Protein Conformation / Protein Domains / Proteins / Science / Science (multidisciplinary) / Translocation