Recruitment of MKLP1 to the spindle midzone/midbody by INCENP is essential for midbody formation and completion of cytokinesis in human cells

Abstract

The INCENP (inner centromere protein) is a chromosomal passenger protein that plays multiple roles in regulating mitosis and cytokinesis. The MKLP1 (mitotic kinesin-like protein) is a component of centralspindlin complex that has been implicated in assembly of midzone/midbody during mitosis and is essential for cytokinesis. In the present study, we investigated functions of INCNEP and MKLP1 and their interplay in regulating spindle midzone/midbody formation and cytokinesis in human cells. Immunofluorescence and live-cell imaging analyses have shown that, in addition to multiple chromosome segregation defects, cells that lacked INCENP by RNAi (RNA interference) exhibit abnormal spindle midzone/midbody formation, resulting in formation of binucleated/multinucleated cells. Suppression of MKLP1 expression by siRNA (small interfering RNA) did not cause any abnormality of chromosome segregation and midzone formation, but abrogated midbody formation and completion of cytokinesis. Furthermore, we show that INCENP is required for recruiting MKLP1 to the spindle midzone/midbody. Three-dimensional reconstruction imaging analysis suggests that recruitment of MKLP1 to the midzone/midbody by INCENP is a crucial step for the midbody formation and completion of cytokinesis in mammalian cells.