Importin-β1 functions as a chromatin sensor to position the contractile ring for cytokinesis

Abstract

Cytokinesis, thefinal step of cell division, relies on the ingression of a carefully positioned actomyosin ring. Chromatin-associated Ran-GTP helps position this ring although the mechanism is not clear. We hypothesize that a depleted zone of Ran-GTP between the segregating chromosomes causes importins to become equatorially enriched and promote the recruitment of the scaffold protein anillin. However, the role of importins during anaphase remains unclear. Here, we tested if importins respond to the chromatin-associated Ran-GTP gradient to regulate ring assembly. Using endogenous tagging, live-cell imaging, and optogenetic perturbations, we found that importin-{beta}1 becomes equatorially enriched and is required for cytokinesis in hypotriploid HeLa cells, but not in euploid HCT 116 cells. A predictive model of the Ran-GTP gradient and Ran-free importin-{beta}1, validated experimentally by FLIM-FRET, identified factors that modulate this chromatin sensing pathway including chromatin-to-cell size ratio. Ourfindings suggest that highly aneuploid cancer cells may depend on importin-mediated anillin recruitment, representing a targetable weakness that distinguishes them from diploid cells. Summary blurbImportin enrichment at the equator promotes anillin recruitment during cytokinesis in a ploidy-dependent manner, linking Ran-GTP gradients to cell division. HighlightsO_LIRan-free Importin-{beta}1 becomes enriched between segregating chromosomes in anaphase HeLa cells but not in HCT116 cells. C_LIO_LIImportin-binding is required for anillin localization and function in cytokinesis in HeLa cells but not in HCT 116 cells. C_LIO_LIImportin-{beta}1 disruption at anaphase causes ring oscillation and cytokinesis failure in HeLa cells. C_LIO_LIImportin-{beta}1 is not equatorially enriched and anillin is broader in HeLa cells with a lower chromatin-to-cell-size ratio. C_LIO_LIImportin-{beta}1 is equatorially enriched and anillin is narrower in HCT 116 cells with a higher chromatin-to-cell-size ratio. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=188 HEIGHT=200 SRC="FIGDIR/small/688041v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@1545a59org.highwire.dtl.DTLVardef@f7b8edorg.highwire.dtl.DTLVardef@18df715org.highwire.dtl.DTLVardef@15891d5_HPS_FORMAT_FIGEXP M_FIG C_FIG