Specific localization of serine 19 phosphorylated myosin II during cell locomotion and mitosis of cultured cells

Fumio Matsumura, Shoichiro Ono, Yoshihiko Yamakita, Go Totsukawa, Shigeko Yamashiro

Research output: Contribution to journalArticle

190 Citations (Scopus)

Abstract

Phosphorylation of the regulatory light chain of myosin II (RMLC) at Serine 19 by a specific enzyme, MLC kinase, is believed to control the contractility of actomyosin in smooth muscle and vertebrate nonmuscle cells. To examine how such phosphorylation is regulated in space and time within cells during coordinated cell movements, including cell locomotion and cell division, we generated a phosphorylation-specific antibody. Motile fibroblasts with a polarized cell shape exhibit a bimodal distribution of phosphorylated myosin along the direction of cell movement. The level of myosin phosphorylation is high in an anterior region near membrane ruffles, as well as in a posterior region containing the nucleus, suggesting that the contractility of both ends is involved in cell locomotion. Phosphorylated myosin is also concentrated in cortical microfilament bundles, indicating that cortical filaments are under tension. The enrichment of phosphorylated myosin in the moving edge is shared with an epithelial cell sheet; peripheral microfilament bundles at the leading edge contain a higher level of phosphorylated myosin. On the other hand, the phosphorylation level of circumferential microfilament bundles in cell-cell contacts is low. These observations suggest that peripheral microfilaments at the edge are involved in force production to drive the cell margin forward while micro filaments in cell-cell contacts play a structural role. During cell division, both fibroblastic and epithelial cells exhibit an increased level of myosin phosphorylation upon cytokinesis, which is consistent with our previous biochemical study (Yamakita, Y., S. Yamashiro, and F. Matsumura. 1994. J. Cell Biol. 124:129-137). In the case of the NRK epithelial cells, phosphorylated myosin first appears in the midzones of the separating chromosomes during late anaphase, but apparently before the formation of cleavage furrows, suggesting that phosphorylation of RMLC is an initial signal for cytokinesis.

Original languageEnglish (US)
Pages (from-to)119-129
Number of pages11
JournalJournal of Cell Biology
Volume140
Issue number1
DOIs
StatePublished - Jan 12 1998

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Myosin Type II
Mitosis
Serine
Myosins
Cell Movement
Cultured Cells
Phosphorylation
Actin Cytoskeleton
Cytokinesis
Epithelial Cells
Cell Division
Actomyosin
Anaphase
Cell Shape
Smooth Muscle
Vertebrates
Phosphotransferases
Fibroblasts
Chromosomes
Light

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Matsumura, Fumio ; Ono, Shoichiro ; Yamakita, Yoshihiko ; Totsukawa, Go ; Yamashiro, Shigeko. / Specific localization of serine 19 phosphorylated myosin II during cell locomotion and mitosis of cultured cells. In: Journal of Cell Biology. 1998 ; Vol. 140, No. 1. pp. 119-129.
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abstract = "Phosphorylation of the regulatory light chain of myosin II (RMLC) at Serine 19 by a specific enzyme, MLC kinase, is believed to control the contractility of actomyosin in smooth muscle and vertebrate nonmuscle cells. To examine how such phosphorylation is regulated in space and time within cells during coordinated cell movements, including cell locomotion and cell division, we generated a phosphorylation-specific antibody. Motile fibroblasts with a polarized cell shape exhibit a bimodal distribution of phosphorylated myosin along the direction of cell movement. The level of myosin phosphorylation is high in an anterior region near membrane ruffles, as well as in a posterior region containing the nucleus, suggesting that the contractility of both ends is involved in cell locomotion. Phosphorylated myosin is also concentrated in cortical microfilament bundles, indicating that cortical filaments are under tension. The enrichment of phosphorylated myosin in the moving edge is shared with an epithelial cell sheet; peripheral microfilament bundles at the leading edge contain a higher level of phosphorylated myosin. On the other hand, the phosphorylation level of circumferential microfilament bundles in cell-cell contacts is low. These observations suggest that peripheral microfilaments at the edge are involved in force production to drive the cell margin forward while micro filaments in cell-cell contacts play a structural role. During cell division, both fibroblastic and epithelial cells exhibit an increased level of myosin phosphorylation upon cytokinesis, which is consistent with our previous biochemical study (Yamakita, Y., S. Yamashiro, and F. Matsumura. 1994. J. Cell Biol. 124:129-137). In the case of the NRK epithelial cells, phosphorylated myosin first appears in the midzones of the separating chromosomes during late anaphase, but apparently before the formation of cleavage furrows, suggesting that phosphorylation of RMLC is an initial signal for cytokinesis.",
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Specific localization of serine 19 phosphorylated myosin II during cell locomotion and mitosis of cultured cells. / Matsumura, Fumio; Ono, Shoichiro; Yamakita, Yoshihiko; Totsukawa, Go; Yamashiro, Shigeko.

In: Journal of Cell Biology, Vol. 140, No. 1, 12.01.1998, p. 119-129.

Research output: Contribution to journalArticle

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