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
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Amino Acid Sequence
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Computational Biology
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Cytoskeleton / metabolism
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Cytoskeleton / ultrastructure
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Gene Expression
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Histidine / genetics
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Histidine / metabolism
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Intrinsically Disordered Proteins / chemistry*
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Intrinsically Disordered Proteins / genetics
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Intrinsically Disordered Proteins / metabolism
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Kinesins / chemistry*
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Kinesins / genetics
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Kinesins / metabolism
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Microtubules / metabolism
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Microtubules / ultrastructure
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Mitosis
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Molecular Sequence Data
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Oligopeptides / genetics
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Oligopeptides / metabolism
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Protein Binding
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Protein Structure, Tertiary
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Schizosaccharomyces / genetics*
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Schizosaccharomyces / metabolism
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Schizosaccharomyces / ultrastructure
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Schizosaccharomyces pombe Proteins / chemistry*
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Schizosaccharomyces pombe Proteins / genetics
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Schizosaccharomyces pombe Proteins / metabolism
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Sequence Alignment
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Spindle Apparatus / metabolism*
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Spindle Apparatus / ultrastructure
Substances
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Cut7 protein, S pombe
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His-His-His-His-His-His
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Intrinsically Disordered Proteins
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Oligopeptides
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Recombinant Fusion Proteins
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Schizosaccharomyces pombe Proteins
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Green Fluorescent Proteins
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Histidine
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Kinesins