Molecular properties of the N-terminal extension of the fission yeast kinesin-5, Cut7

Genet Mol Res. 2016 Feb 11;15(1). doi: 10.4238/gmr.15017799.

Abstract

Kinesin-5 plays an essential role in spindle formation and function, and serves as a potential target for anti-cancer drugs. The aim of this study was to elucidate the molecular properties of the N-terminal extension of the Schizosaccharomyces pombe kinesin-5, Cut7. This extension is rich in charged amino acids and predicted to be intrinsically disordered. In S. pombe cells, a Cut7 construct lacking half the N-terminal extension failed to localize along the spindle microtubules and formed a monopolar spindle. However, a construct lacking the entire N-terminal extension exhibited normal localization and formed a typical bipolar spindle. In addition, in vitro analyses revealed that the truncated Cut7 constructs demonstrated similar motile velocities and directionalities as the wild-type motor protein, but the microtubule landing rates were significantly reduced. These findings suggest that the N-terminal extension is not required for normal Cut7 intracellular localization or function, but alters the microtubule-binding properties of this protein in vitro.

MeSH terms

  • Amino Acid Sequence
  • Computational Biology
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure
  • Gene Expression
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Histidine / genetics
  • Histidine / metabolism
  • Intrinsically Disordered Proteins / chemistry*
  • Intrinsically Disordered Proteins / genetics
  • Intrinsically Disordered Proteins / metabolism
  • Kinesins / chemistry*
  • Kinesins / genetics
  • Kinesins / metabolism
  • Microtubules / metabolism
  • Microtubules / ultrastructure
  • Mitosis
  • Molecular Sequence Data
  • Oligopeptides / genetics
  • Oligopeptides / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces / ultrastructure
  • Schizosaccharomyces pombe Proteins / chemistry*
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism
  • Sequence Alignment
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / ultrastructure

Substances

  • Cut7 protein, S pombe
  • His-His-His-His-His-His
  • Intrinsically Disordered Proteins
  • Oligopeptides
  • Recombinant Fusion Proteins
  • Schizosaccharomyces pombe Proteins
  • Green Fluorescent Proteins
  • Histidine
  • Kinesins