Mutations in the bimC box of Cut7 indicate divergence of regulation within the bimC family of kinesin related proteins.
Members of the bimC family of kinesin related proteins (KRPs) play vital roles in the formation and function of the mitotic spindle. Although they share little amino acid homology outside the highly conserved microtubule motor domain, several family members do contain a 'bimC box', a sequence motif around a p34(cdc2) consensus phosphorylation site in their carboxy-terminal 'tail' region. One family member, Eg5, requires phosphorylation at this site for association with the mitotic spindle. We show that mutations in the Schizosaccharomyces pombe cut7+ gene that change the bimC box p34(cdc2) consensus phosphorylation site at position 1,011 and a neighbouring MAP kinase consensus phosphorylation site at position 1,020 to non-phosphorylatable residues did not affect the ability of S. pombe cut7 genes to complement temperature sensitive cut7 mutants. Phosphorylation site mutants expressed as fusions to green fluorescent protein associated with the mitotic spindle with a localisation indistinguishable from similarly expressed wild-type Cut7. Cells in which cut7.T1011A replaced the genomic copy of cut7+ were viable and formed normal spindles. Deletion of the entire carboxy-terminal tail region did not affect the ability of Cut7 to associate with the mitotic spindle but did inhibit normal spindle formation. Thus, unlike Eg5, neither the p34(cdc2) consensus phosphorylation site in the bimC box nor the entire tail region of Cut7 are required for association with the mitotic spindle.
Drummond
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DR
School of Biological Sciences, University of Manchester, Manchester, UK.
Hagan
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D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D018628:Gene Dosage; D015966:Gene Expression Regulation, Fungal; D006801:Humans; D016547:Kinesins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins
C094560:BimC protein, Aspergillus nidulans; C104159:Cut7 protein, S pombe; D005656:Fungal Proteins; D029702:Schizosaccharomyces pombe Proteins; D016547:Kinesins