Function of a minus-end-directed kinesin-like motor protein in mammalian cells

Jurgita Matuliene, Russell Essner, Jung Hwa Ryu, Yukihisa Hamaguchi, Peter W. Baas, Tokuko Haraguchi, Yasushi Hiraoka, Ryoko Kuriyama

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

CHO2 is a mammalian minus-end-directed kinesin-like motor protein present in interphase centrosomes/nuclei and mitotic spindle fibers/poles. Expression of HA- or GFP-tagged subfragments in transfected CHO cells revealed the presence of the nuclear localization site at the N-terminal tail. This domain becomes associated with spindle fibers during mitosis, indicating that the tail is capable of interaction with microtubules in vivo. While the central stalk diffusely distributes in the entire cytoplasm of cells, the motor domain co-localizes with microtubules throughout the cell cycle, which is eliminated by mutation of the ATP-binding consensus motif from GKT to AAA. Overexpression of the full-length CHO2 causes mitotic arrest and spindle abnormality. The effect of protein expression was first seen around the polar region where microtubule tended to be bundled together. A higher level of protein expression induces more elongated spindles which eventually become disorganized by loosing the structural integrity between microtubule bundles. Live cell observation demonstrated that GFP-labeled microtubule bundles underwent continuous changes in their relative position to one another through repeated attachment and detachment at one end; this results in the formation of irregular number of microtubule focal points in mitotic arrested cells. Thus the primary action of CHO2 appears to cross-link microtubules and move toward the minus-end direction to maintain association of the microtubule end at the pole. In contrast to the full-length of CHO2, overexpression of neither truncated nor mutant polypeptides resulted in significant effects on mitosis and mitotic spindles, suggesting that the function of CHO2 in mammalian cells may be redundant with other motor molecules during cell division.

Original languageEnglish (US)
Pages (from-to)4041-4050
Number of pages10
JournalJournal of cell science
Volume112
Issue number22
StatePublished - Dec 1 1999

Keywords

  • CHO cell
  • Microtubule
  • Minus-end motor
  • Mitotic spindle

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