Blikstad, Ingrid and Carlsson, Lars (1982) On the dynamics of the microfilament system in HeLa cells. Journal of Cell Biology, 93 (1). pp. 122-128. ISSN 0021-9525 http://resolver.caltech.edu/CaltechAUTHORS:BLIjbc82
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We measured the pools of unpolymerized and filamentous actin in homogenates of HeLa 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 a series of 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 percent of the total actin in an unpolymerized form. With some batches of cells this number can be as high as 50 percent. In sparse cultures (3 x 10(4) cell/cm(2)), HeLa cells contain approximately 10 pg actin/cell, while the corresponding number is only 5 pg in dense cultures (3 x 10(5) cells/cm(2)). Treatment of cells with cytochalasin B increases the pool of unpolymerized actin by approximately 30-40 percent, while colchicine decreases the fraction of unpolymerized actin by 20 percent. The oxidant diamide increases the filamentous actin pool 25-50 percent. 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. These results show that significant changes in the actin pools can be induced in nucleated cells. However, several treatments which alter the morphology and motility of cells are not accompanied by an alteration in the G-/F-actin ratio.
|Additional Information:||Copyright © 1982 by The Rockefeller University Press. Received for publication 26 May 1981, and in revised form 13 October 1981. We thank Jan Stenlid for help with culturing of cells during an intense period in this work. We also express our gratitude to Anna-Greta Lundquist and Solveig Mill for excellent secretarial work. This investigation was financially supported by grants to the authors from P. E. Lindahls Foundation, Lennander Foundation, and Jeanssons Foundation, and grants to Professor Uno Lindberg from Uppsala University Reserve Foundation, the Knut and Alice Wallenberg Foundation, and the Swedish Cancer Society, which we gratefully acknowledge, L. Carlsson is a recipient of a Research fellowship from the Swedish Natural Science Research Council.|
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|Deposited By:||Lindsay Cleary|
|Deposited On:||23 Aug 2006|
|Last Modified:||26 Dec 2012 08:59|
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