The yeast dynactin complex is involved in partitioning the mitotic spindle between mother and daughter cells during anaphase B.
Although vertebrate cytoplasmic dynein can move to the minus ends of microtubules in vitro, its ability to translocate purified vesicles on microtubules depends on the presence of an accessory complex known as dynactin. We have cloned and characterized a novel gene, NIP100, which encodes the yeast homologue of the vertebrate dynactin complex protein p150(glued). Like strains lacking the cytoplasmic dynein heavy chain Dyn1p or the centractin homologue Act5p, nip100Delta strains are viable but undergo a significant number of failed mitoses in which the mitotic spindle does not properly partition into the daughter cell. Analysis of spindle dynamics by time-lapse digital microscopy indicates that the precise role of Nip100p during anaphase is to promote the translocation of the partially elongated mitotic spindle through the bud neck. Consistent with the presence of a true dynactin complex in yeast, Nip100p exists in a stable complex with Act5p as well as Jnm1p, another protein required for proper spindle partitioning during anaphase. Moreover, genetic depletion experiments indicate that the binding of Nip100p to Act5p is dependent on the presence of Jnm1p. Finally, we find that a fusion of Nip100p to the green fluorescent protein localizes to the spindle poles throughout the cell cycle. Taken together, these results suggest that the yeast dynactin complex and cytoplasmic dynein together define a physiological pathway that is responsible for spindle translocation late in anaphase.
Kahana
J A
JA
Department of Cancer Biology, Dana-Farber Cancer Institute, and Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Schlenstedt
G
G
Evanchuk
D M
DM
Geiser
J R
JR
Hoyt
M A
MA
Silver
P A
PA
D000595:Amino Acid Sequence; D000705:Anaphase / Q000235:genetics / Q000502:physiology*; D000818:Animals; D000072159:Dynactin Complex; D004398:Dyneins / Q000737:chemistry / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D008969:Molecular Sequence Data; D051381:Rats; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000235:genetics / Q000502:physiology*
D000072159:Dynactin Complex; D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; C000606366:NIP100 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D004398:Dyneins