The actomyosin network is influenced by NMHC IIA and regulated by CrpF46, which is involved in controlling cell migration

Abstract When a cell migrates, the centrosome positions between the nucleus and the leading edge of migration via the microtubule system. The protein Crp F46 (centrosome-related protein F46) has a known role during mitosis and centrosome duplication. However, how Crp F46 efficiently regulates centrosome-related cell migration is unclear. Here, we report that knockdown of Crp F46 resulted in the disruption of microtubule arrangement, with impaired centrosomal reorientation, and slowed down cell migration. In cells that express low levels of Crp F46 , stress fibers were weakened, which could be rescued by recovering Flag-Crp F46 . We also found that Crp F46 interacted with non-muscle myosin high chain IIA (NMHC IIA) and that its three coiled-coil domains are pivotal for its binding to NMHC IIA. Additionally, analyses of phosphorylation of NMHC IIA and RLC (regulatory light chain) demonstrated that Crp F46 was associated with myosin IIA during filament formation. Indirect immunofluorescence images indicated that NM IIA filaments were inhibited when Crp F46 was under-expressed. Thus, Crp F46 regulates cell migration by centrosomal reorientation and altering the function of the actomyosin network by controlling specific phosphorylation of myosin.