Multiple mechanisms of chromosome movement in vertebrate cells mediated through the Ndc80 complex and dynein/dynactin.
Kinetochores bind microtubules laterally in a transient fashion and stably, by insertion of plus ends. These pathways may exist to carry out distinct tasks during different stages of mitosis and likely depend on distinct molecular mechanisms. On isolated chromosomes, we found microtubule nucleation/binding depended additively on both dynein/dynactin and on the Ndc80/Hec1 complex. Studying chromosome movement in living Xenopus cells within the simplified geometry of monopolar spindles, we quantified the relative contributions of dynein/dynactin and the Ndc80/Hec1 complex. Inhibition of dynein/dynactin alone had minor effects but did suppress transient, rapid, poleward movements. In contrast, inhibition of the Ndc80 complex blocked normal end-on attachments of microtubules to kinetochores resulting in persistent rapid poleward movements that required dynein/dynactin. In normal cells with bipolar spindles, dynein/dynactin activity on its own allowed attachment and rapid movement of chromosomes on prometaphase spindles but failed to support metaphase alignment and chromatid movement in anaphase. Thus, in prometaphase, dynein/dynactin likely mediates early transient, lateral interactions of kinetochores and microtubules. However, mature attachment via the Ndc80 complex is essential for metaphase alignment and anaphase A.
Molecular, Cell and Developmental Biology Research Program, Oklahoma Medical Research Foundation, 825 NE 13th St, MS 48, Oklahoma City, OK 73104, USA.
Valeriya V
VV
Vorozhko
Michael J
MJ
Emanuele
Marko J
MJ
Kallio
P Todd
PT
Stukenberg
Gary J
GJ
Gorbsky
D000705:Anaphase; D000818:Animals; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002875:Chromosomes / Q000378:metabolism / Q000648:ultrastructure*; D003598:Cytoskeletal Proteins; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D008954:Models, Biological; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008941:Spindle Apparatus; D014162:Transfection; D014404:Tubulin / Q000378:metabolism; D014982:Xenopus laevis
D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; C120529:NDC80 protein, human; C530471:Ndc80 protein, Xenopus; D009687:Nuclear Proteins; D014404:Tubulin; D004398:Dyneins