On the dynamics of the microfilament system in HeLa cells

Abstract

Pools of unpolymerized and filamentous actin were measured in homogenates of HeLa [human cervical carcinoma] cells made in several different lysis buffers, as well as after treatment of cells with a variety of chemicals or trypsin, and after adenovirus (type 2) infection. This was possible when certain factors concerning the basic culture conditions were kept constant: e.g., serum type used, serum batch, cell density, time after subcultivation of cells and buffering substance in the medium. Homogenates from untreated cells usually contain 35-45% of the total actin in an unpolymerized from. With some batches of cells this number can be as high as 50%. In sparse culture (3 .times. 104 cells/cm2), HeLa cells contain .apprx. 10 pg actin/cell, while the corresponding number is only 5 pg in dense cultures (3 .times. 105 cells/cm2). Treatment of cells with cytochalasin B increases the pool of unpolymerized actin by .apprx. 80-40%, while colchicine decreases the fraction of unpolymerized actin by 20%. The oxidant diamide increases the filamentous actin pool 25-50%. Glucose, sodium azide, dinitrophenol, serum starvation or thymidine treatment does not affect the distribution between unpolymerized and filamentous actin to any significant extent. Trypsin and EDTA induced rounding up of cells but did not change the actin distribution. The distribution of actin between G- and F-forms was unchanged after adenovirus infection. Significant changes in the actin pools can be induced in nucleated cells. Several treatments which alter the morphology and motility of cells are not accompanied by an alteration in the G-/F-actin rtio.