M. Novakova et al., GAMMA-TUBULIN REDISTRIBUTION IN TAXOL-TREATED MITOTIC CELLS PROBED BYMONOCLONAL-ANTIBODIES, Cell motility and the cytoskeleton, 33(1), 1996, pp. 38-51
Monoclonal antibodies were prepared against conserved synthetic peptid
e from the C-terminus of the gamma-tubulin and their specificity was c
onfirmed by immunoblotting, competitive enzyme-linked immunosorbent as
say (ELISA) and immunofluorescence. The antibodies decorated interphas
e centrosomes as well as half-spindles and midbodies in mitotic cells
of various origin. The prepared antibodies were used to study the gamm
a-tubulin distribution in nocodazole and taxol-treated cells. In the c
ells recovering from the nocodazole treatment, gamma-tubulin was found
in centers of all microtubule asters. Examination of relative locatio
n of gamma-tubulin and microtubule asters in taxol-treated mitotic cel
ls 3T3, HeLa and PtK2 revealed that the number of taxol-induced microt
ubule asters exceeded the number of gamma-tubulin-positive spots. The
gamma-tubulin was often found in the periphery of microtubule asters.
Centrosomal phosphoprotein epitope detected by MPM-2 antibody colocali
zed with gamma-tubulin in taxol-treated mitotic cells. The presented d
ata suggest that taxol-induced microtubule asters are in vivo nucleate
d independently of gamma-tubulin, and other minus-end nucleator(s) are
necessary for formation of such asters. Alternatively, gamma-tubulin
is present in subthreshold amounts undetectable by immunofluorescence.
(C) 1996 Wiley-Liss, Inc.