PRC1 is a microtubule binding and bundling protein essential to maintain the mitotic spindle midzone.
Midzone microtubules of mammalian cells play an essential role in the induction of cell cleavage, serving as a platform for a number of proteins that play a part in cytokinesis. We demonstrate that PRC1, a mitotic spindle-associated Cdk substrate that is essential to cell cleavage, is a microtubule binding and bundling protein both in vivo and in vitro. Overexpression of PRC1 extensively bundles interphase microtubules, but does not affect early mitotic spindle organization. PRC1 contains two Cdk phosphorylation motifs, and phosphorylation is possibly important to mitotic suppression of bundling, as a Cdk phosphorylation-null mutant causes extensive bundling of the prometaphase spindle. Complete suppression of PRC1 by siRNA causes failure of microtubule interdigitation between half spindles and the absence of a spindle midzone. Truncation mutants demonstrate that the NH2-terminal region of PRC1, rich in alpha-helical sequence, is important for localization to the cleavage furrow and to the center of the midbody, whereas the central region, with the highest sequence homology between species, is required for microtubule binding and bundling activity. We conclude that PRC1 is a microtubule-associated protein required to maintain the spindle midzone, and that distinct functions are associated with modular elements of the primary sequence.
Mollinari
Cristiana
C
Institut de Biologie Structurale J-P Ebel (CEA-CNRS), 38027 Grenoble cedex 1, France.
Kleman
Jean-Philippe
JP
Jiang
Wei
W
Schoehn
Guy
G
Hunter
Tony
T
Margolis
Robert L
RL
D020816:Amino Acid Motifs; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D018385:Centrosome / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000737:chemistry; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D007399:Interphase; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D009154:Mutation; D017239:Paclitaxel / Q000494:pharmacology; D010766:Phosphorylation; D034741:RNA, Small Interfering; D022661:RNA, Untranslated / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000502:physiology*; D014407:Tumor Cells, Cultured
D018797:Cell Cycle Proteins; D006657:Histones; C116662:PRC1 protein, human; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D011993:Recombinant Fusion Proteins; D018844:Cyclin-Dependent Kinases; D017239:Paclitaxel