Regulation of cytoskeletal mechanics and cell growth by myosin light chain phosphorylation

Shuang Cai, Lidija Pestic-Dragovich, Martha E. O'Donnell, Ning Wang, Donald Ingber, Elliot Elson, Primal De Lanerolle

Research output: Contribution to journalArticlepeer-review

Abstract

The role of myosin light chain phosphorylation in regulating the mechanical properties of the cytoskeleton was studied in NIH/3T3 fibroblasts expressing a truncated, constitutively active form of smooth muscle myosin light chain kinase (tMK). Cytoskeletal stiffness determined by quantifying the force required to indent the apical surface of adherent cells showed that stiffness was increased twofold in tMK cells compared with control cells expressing the empty plasmid (Neo cells). Cytoskeletal stiffness quantified using magnetic twisting cytometry showed an ~1.5-fold increase in stiffness in tMK cells compared with Neo cells. Electronic volume measurements on cells in suspension revealed that tMK cells had a smaller volume and are more resistant to osmotic swelling than Neo cells. tMK cells also have smaller nuclei, and activation of mitogen-activated protein kinase (MAP kinase) and translocation of MAP kinase to the nucleus are slower in tMK cells than in control cells. In tMK cells, there is also less bromodeoxyuridine incorporation, and the doubling time is increased. These data demonstrate that increased myosin light chain phosphorylation correlates with increased cytoskeletal stiffness and suggest that changing the mechanical characteristics of the cytoskeleton alters the intracellular signaling pathways that regulate cell growth and division.

Original languageEnglish (US)
Pages (from-to)C1349-C1356
JournalAmerican Journal of Physiology - Cell Physiology
Volume275
Issue number5 44-5
DOIs
StatePublished - 1998
Externally publishedYes

Keywords

  • Cell division
  • Cell stiffness
  • Mitogen- activated protein kinase activation
  • Myosin light chain kinase
  • Osmotic swelling
  • Volume regulation

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

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