Protein synthesis requires cell-surface contact while nuclear events respond to cell shape in anchorage-dependent fibroblasts

Abstract

Anchorage-dependent mouse fibroblasts grow only if attached to and spread on a solid substrate. The suspension of cells in methyl-cellulose results in dramatic, coordinated inhibition of the major RNA and protein synthesis systems, and these systems are sequentially restored when cells are replated on a tissue culture dish surface. In the present report the effects on metabolism of cell reattachment are separated from those of subsequent spreading by controlling cell shape. Macromolecular metabolism is first strongly suppressed by long-term suspension culture. The cells are then replated in the presence of a variety of spreading inhibitors. The recovery of protein synthesis, which rapidly follows reattachment, does not require extensive cell spreading. Contact of a limited portion of the plasma membrane with the solid culture dish surface is apparently a sufficient signal by itself. A very different method of controlling cell shape is afforded by changing culture dish surface adhesivity. Suspended cells are replated on dishes precoated with thin layers of the hydrophilic hydrogen poly(2-hydroxyethyl methacrylate). The final mean cell diameter is then varied over wide limits. As before, protein synthesis recovery is unaffected. However, nuclear events such as DNA and rRNA synthesis and mRNA production are profoundly affected by cell shape. Thus, cell surface contact and cell shape give rise to distinctly different regulatory responses.