Roles of kinesin and kinesin-like proteins in sea urchin embryonic cell division: evaluation using antibody microinjection.
Previous studies suggest that kinesin heavy chain (KHC) is associated with ER-derived membranes that accumulate in the mitotic apparatus in cells of early sea urchin embryos (Wright, B. D., J. H. Henson, K. P. Wedaman, P. J. Willy, J. N. Morand, and J. M. Scholey. 1991. J. Cell Biol. 113:817-833). Here, we report that the microinjection of KHC-specific antibodies into these cells has no effect on mitosis or ER membrane organization, even though one such antibody, SUK4, blocks kinesin-driven motility in vitro and in mammalian cells. Microinjected SUK4 was localized to early mitotic figures, suggesting that it is able to access kinesin in spindles. In contrast to KHC-specific antibodies, two antibodies that react with kinesin-like proteins (KLPs), namely CHO1 and HD, disrupted mitosis and prevented subsequent cell division. CHO1 is thought to exert this effect by blocking the activity of a 110-kD KLP. The relevant target of HD, which was raised against the KHC motor domain, is unknown; HD may disrupt mitosis by interfering with an essential spindle KLP but not with KHC itself, as preabsorption of HD with KHC did not alter its ability to block mitosis. These data indicate that some KLPs have essential mitotic functions in early sea urchin embryos but KHC itself does not.
Wright
B D
BD
Section of Molecular and Cellular Biology, University of California at Davis 95616.
Terasaki
M
M
Scholey
J M
JM
D000818:Animals; D000906:Antibodies / Q000008:administration & dosage / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology*; D002465:Cell Movement / Q000187:drug effects; D004625:Embryo, Nonmammalian / Q000166:cytology* / Q000378:metabolism; D005260:Female; D005306:Fertilization; D016547:Kinesins / Q000276:immunology / Q000502:physiology*; D008297:Male; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D012617:Sea Urchins / Q000196:embryology*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D016547:Kinesins