Phosphorylation of CHO1 by Lats1/2 regulates the centrosomal activation of LIMK1 during cytokinesis

Abstract

Large tumor suppressor 1 and 2 (Lats1/2) regulate centrosomal integrity, chromosome segregation and cytokinesis. As components of the centralspindlin complex, the kinesin-like protein CHO1 and its splicing variant MKLP1 colocalize with chromosome passenger proteins and GTPases and regulate the formation of the contractile ring and cytokinesis; however, the regulatory mechanisms of CHO1/MKLP1 remain elusive. Here, we show that Lats1/2 phosphorylate Ser716 in the F-actin-interacting region of CHO1, which is absent in MKLP1. Phosphorylated CHO1 localized to the centrosomes and midbody, and the actin polymerization factor LIM-kinase 1 (LIMK1) was identified as its binding partner. Overexpression of constitutively phosphorylated and non-phosphorylated CHO1 altered the mitotic localization and activation of LIMK1 at the centrosomes in HeLa cells, leading to the inhibition of cytokinesis through excessive phosphorylation of Cofilin and mislocalization of Ect2. These results suggest that Lats1/2 stringently control cytokinesis by regulating CHO1 phosphorylation and the mitotic activation of LIMK1 on centrosomes.

Hippo signaling is intrinsically regulated during cell cycle progression by APC/C Cdh1

“…Importantly, LATS1 regulates actin polymerization that affects cytokinesis through negative modulation of LIMK1 (65). Indeed, there is evidence that LATS1/2 stringently control cytokinesis by regulating CHO1 phosphorylation and mitotic activation of LIMK1 on centrosomes (66). Finally, LATS1 has been suggested to be a component of the mitotic exit network in higher eukaryotes (67), which may explain its abilities to induce G2 arrest and regulate cytokinesis (68)(69)(70).…”

Large tumor suppressors 1 and 2 regulate Aurora-B through phosphorylation of INCENP to ensure completion of cytokinesis

“…HeLa-S3 (human cervical carcinoma) and HEK293T (human embryonic kidney) cells were cultured in DMEM (Sigma, St. Louis, MO, USA) with 5% or 10% FBS (Hyclone, Logan, UT, USA) supplemented with 100 U/ml penicillin and 100 μg/ml streptomycin. TALEN-mediated LATS2 -knockout HeLa-S3 cell lines (LATS2-KO/HeLa-S3) were generated and maintained in DMEM with 5% FBS, as described previously ( Okamoto et al, 2015 ). Cell-cycle synchronization was performed as described previously ( Yabuta et al, 2011 ), except as specifically noted.…”

“…LATS1 and LATS2 (LATS1/2) are also directly involved in mitotic regulation. For example, in mammalian cells, dysregulation of particular sub-pathways of LATS-mediated signaling leads to severe defects in mitotic progression, mitotic exit, and cytokinesis ( Yang et al, 2004 ; Bothos et al, 2005 ; Yabuta et al, 2007 ; Yabuta et al, 2011 ; Chiyoda et al, 2012 , Okamoto et al, 2015 ). Moreover, LATS2 also regulates a cell-cycle checkpoint to prevent polyploidization via p53 and Yap in response to centrosome damage ( Aylon et al, 2006 ; Ganem et al, 2014 ).…”

The Hippo Signaling Pathway in Cancer: A Cell Cycle Perspective

“…Mechanistically, the LATS1/2-mediated phosphorylation of INCENP at S894 is involved in Aurora B activation, telophase progression, and cytokinesis [ 133 ]. One study demonstrates that LATS1/2 phosphorylates CHO1 at S716, which promotes the binding and activation of LIMK1 at centrosome to trigger cytokinesis initiation [ 136 ]. However, another study indicates that LATS1 inhibits the kinase activity of LIMK1 at the contractile ring to control cytokinesis [ 137 ].…”