title: Enhancement of myosin II/actin turnover at the contractile ring induces slower furrowing in dividing HeLa cells.
abstract: Myosin II ATPase activity is enhanced by the phosphorylation of MRLC (myosin II regulatory light chain) in non-muscle cells. It is well known that pMRLC (phosphorylated MRLC) co-localizes with F-actin (filamentous actin) in the CR (contractile ring) of dividing cells. Recently, we reported that HeLa cells expressing non-phosphorylatable MRLC show a delay in the speed of furrow ingression, suggesting that pMRLC plays an important role in the control of furrow ingression. However, it is still unclear how pMRLC regulates myosin II and F-actin at the CR to control furrow ingression during cytokinesis. In the present study, to clarify the roles of pMRLC, we measured the turnover of myosin II and actin at the CR in dividing HeLa cells expressing fluorescent-tagged MRLCs and actin by FRAP (fluorescence recovery after photobleaching). A myosin II inhibitor, blebbistatin, caused an enhancement of the turnover of MRLC and actin at the CR, which induced a delay in furrow ingression. Furthermore, only non-phosphorylatable MRLC and a Rho-kinase inhibitor, Y-27632, accelerated the turnover of MRLC and actin at the CR. Interestingly, the effect of Y-27632 was cancelled in the cell expressing phosphomimic MRLCs. Taken together, these results reveal that pMRLC reduces the turnover of myosin II and also actin at the CR. In conclusion, we show that the enhancement of myosin II and actin turnover at the CR induced slower furrowing in dividing HeLa cells.
authors.affiliation: Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, 739-8526, Japan.
authors.forename: Tomo
authors.initials: T
authors.lastname: Kondo
authors.forename: Kozue
authors.initials: K
authors.lastname: Hamao
authors.forename: Keiju
authors.initials: K
authors.lastname: Kamijo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.forename: Makiko
authors.initials: M
authors.lastname: Morita
authors.forename: Masayuki
authors.initials: M
authors.lastname: Takahashi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Hosoya
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000577:Amides; D002462:Cell Membrane / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology; D006367:HeLa Cells; D006576:Heterocyclic Compounds, 4 or More Rings; D006801:Humans; D008938:Mitosis / Q000502:physiology*; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D011725:Pyridines
chemical_list: D000199:Actins; D000577:Amides; D006576:Heterocyclic Compounds, 4 or More Rings; D018994:Myosin Light Chains; D011725:Pyridines; C108830:Y 27632; C472645:blebbistatin; D024462:Myosin Type II