Motor activity and mitotic spindle localization of the Drosophila kinesin-like protein KLP61F.
The KLP61F gene product is essential for Drosophila development. Mutations in KLP61F display a mitotic arrest phenotype caused by a failure in the proper separation of duplicated centrosomes (Heck et al., 1993). Sequence analysis of KLP61F identified it as a member of the bimC family of kinesin-like microtubule motor proteins. Here we report that KLP61F is distinct from KRP130, a kinesin-like protein recently purified from Drosophila embryos and suggested to be the product of the KLP61F gene (Cole et al., 1994). We also characterized recombinant KLP61F and found that it possesses microtubule-stimulated ATPase and microtubule translocation activities in vitro. In addition, we have used an affinity-purified, KLP61F-specific antiserum to localize native KLP61F and an epitope-tagged KLP61F fusion protein during various stages of mitosis in Drosophila syncytial blastoderm embryos. From early prophase through anaphase, KLP61F is coincident with the distribution of tubulin. Together these results confirm the existence of multiple bimC-like kinesins in Drosophila and suggest that KLP61F function is intrinsic to the mitotic spindle.
Barton
N R
NR
Howard Hughes Medical Institute, University of California San Diego, Department of Pharmacology, La Jolla 92093-0683, USA.
Pereira
A J
AJ
Goldstein
L S
LS
D000251:Adenosine Triphosphatases / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D004330:Drosophila / Q000196:embryology / Q000235:genetics / Q000502:physiology*; D029721:Drosophila Proteins; D004625:Embryo, Nonmammalian / Q000502:physiology; D000939:Epitopes; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D017344:Genes, Insect; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000096:biosynthesis / Q000502:physiology*; D008870:Microtubules / Q000648:ultrastructure; D016296:Mutagenesis; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D016324:Sequence Tagged Sites; D008941:Spindle Apparatus / Q000502:physiology* / Q000648:ultrastructure
D029721:Drosophila Proteins; D000939:Epitopes; C084034:Klp61F protein, Drosophila; D008869:Microtubule-Associated Proteins; D011993:Recombinant Fusion Proteins; D000251:Adenosine Triphosphatases