Cj. Hogan et al., INHIBITION OF ANAPHASE SPINDLE ELONGATION IN-VITRO BY A PEPTIDE ANTIBODY THAT RECOGNIZES KINESIN MOTOR DOMAIN, Proceedings of the National Academy of Sciences of the United Statesof America, 90(14), 1993, pp. 6611-6615
Isolated central spindles or spindles in detergent-permeabilized cells
from the diatom Cylindrotheca fusiformis can undergo ATP-dependent re
activation of spindle elongation in vitro. We have used a peptide anti
body raised against a 10-amino add portion common to the kinesin super
family motor domain to look for kinesin-like motor activity during ana
phase B of mitosis. The peptide antibody localizes to central spindles
. Upon ATP reactivation of spindle elongation, antigens recognized by
the antibody are associated exclusively with the central spindle midzo
ne where antiparallel microtubules of each half-spindle overlap. The a
ntibody recognizes several polypeptides by immunoblot using isolated s
pindle extracts. One of these polypeptides behaves like kinesin with r
espect to nucleotide-specific binding to and release from taxol-stabil
ized microtubules. Preincubation of the spindle model with the peptide
antibody inhibits subsequent ATP reactivation of spindle elongation.
Coincubation of the peptide antibody with peptide antigen rescues spin
dle function. These results support a role for kinesin-related protein
(s) in spindle elongation (anaphase B) of mitosis and suggest that one
or several polypeptides that we have identified in spindle extracts m
ay fulfill this function.