Dynamic regulation of ubiquitylation and deubiquitylation at the central spindle during cytokinesis.
During cytokinesis, the central spindle, a bundle of interdigitated anti-parallel microtubules between separating chromosomes, recruits various cytokinetic regulator proteins to the cleavage region. Here, we show that the level of protein ubiquitylation is strikingly and transiently elevated in Aurora B kinase-positive double-band regions of the central spindle during cytokinesis. Two deubiquitylating enzymes UBPY and AMSH, which act on endosomes in interphase, were also recruited to the cleavage region. Whereas UBPY was detected only in the final stage of cytokinesis at the midbody, AMSH localized to a ring structure surrounding the mitotic kinesin MKLP1-positive region of the central spindle and midbody throughout cytokinesis. Depletion of cellular UBPY or AMSH led to defects in cytokinesis. VAMP8, a v-SNARE required for vesicle fusion in cytokinesis, localized to the central spindle region positive for ubiquitylated proteins, and underwent ubiquitylation and deubiquitylation by both UBPY and AMSH. Our results thus implicate the ubiquitylation/deubiquitylation of proteins including VAMP8 in cytokinesis.
Mukai
Akiko
A
Department of Biological Sciences, Tokyo Institute of Technology, 4259-B-16 Nagatsuta, Yokohama 226-8501, Japan.
Mizuno
Emi
E
Kobayashi
Kaoru
K
Matsumoto
Masaki
M
Nakayama
Keiichi I
KI
Kitamura
Naomi
N
Komada
Masayuki
M
D001483:Base Sequence; D002453:Cell Cycle; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference; D008941:Spindle Apparatus; D013347:Subcellular Fractions / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism*
D017931:DNA Primers; D025801:Ubiquitin