Cdc2-mediated phosphorylation of Kid controls its distribution to spindle and chromosomes.
The chromokinesin Kid is important in chromosome alignment at the metaphase plate. Here, we report that Kid function is regulated by phosphorylation. We identify Ser427 and Thr463 as M phase-specific phosphorylation sites and Cdc2-cyclin B as a Thr463 kinase. Kid with a Thr463 to alanine mutation fails to be localized on chromosomes and is only detected along spindles, although it retains the ability to bind DNA or chromosomes. Localization of rigor-type mutant Kid, which shows nucleotide-independent microtubule association, is also confined to the spindle, implying that strong association of Kid with the spindle can sequester it from chromosomes. T463A substitution in DNA-binding domain-truncated Kid consistently enhances its spindle localization. At physiological ionic strength, unphosphorylated Kid shows ATP-independent microtubule association, whereas Thr463-phosphorylated Kid shows ATP dependency. Moreover, the stalk region of unphosphorylated Kid interacts with microtubules and the interaction is weakened when Thr463 is phosphorylated. Our data suggest that phosphorylation on Thr463 of Kid downregulates its affinity for microtubules to ensure reversible association with spindles, allowing Kid to bind chromosomes and exhibit its function.
Departments of Oncology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokane-dai, Minato-ku, Tokyo 108-8639, Japan.
Miho
M
Ohsugi
Noriko
N
Tokai-Nishizumi
Katsuyuki
K
Shiroguchi
Yoko Y
YY
Toyoshima
Jun-Ichiro
J
Inoue
Tadashi
T
Yamamoto
D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002874:Chromosome Mapping; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D017931:DNA Primers; C068353:Kid protein, Bacteria; D013912:Threonine; D012694:Serine; D016203:CDC2 Protein Kinase