Moon, Randall T. and Lazarides, Elias (1983) Canavanine Inhibits Vimentin Assembly But Not Its Synthesis in Chicken Embryo Erythroid Cells. Journal of Cell Biology, 97 (4). pp. 1309-1314. ISSN 0021-9525 http://resolver.caltech.edu/CaltechAUTHORS:20120717-090743486
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In chicken embryo erythroid cells, newly synthesized vimentin first enters a Triton X-100 (TX-100)-soluble pool and subsequently assembles posttranslationally into TX-100-insoluble vimentin filaments (Blikstad I., and E. Lazarides, J. Cell Biol., 96:1803-1808). Here we show that incubation of chicken embryo erythroid cells in a medium in which arginine has been substituted by its amino acid analogue, canavanine, results in the inhibition of the posttranslational assembly of vimentin into the TX-100-insoluble filaments. Immunoprecipitation and subsequent SDS gel electrophoresis showed that the synthesis of canavanine-vimentin is not inhibited and that it accumulates in the TX-100-soluble compartment. Pulse-chase experiments with [35S]methionine demonstrated that while arginine-vimentin can be rapidly chased from the soluble to the cytoskeletal fraction, canavanine-vimentin remains in the soluble fraction, where it turns over. The effect of canavanine on the assembly of vimentin did not prevent the assembly of arginine-vimentin, as cells labeled with [35S]methionine first in the presence of canavanine and then in the presence of arginine contained labeled canavanine-vimentin only in the soluble fraction, and arginine-vimentin in both the soluble and cytoskeletal fractions. These results suggest that arginine residues play an essential role in the assembly of vimentin in vivo.
|Additional Information:||© 1983 Rockefeller University Press. Received for publication 31 May 1983, and in revised form 26 July 1983. We thank J. Ngai, and Drs. W. J. Nelson and B. L. Granger for comments on the manuscript, and W. J. Nelson for communication of work in press. This work was supported by grants from the National Institutes of Health, the National Science Foundation, and the Muscular Dystrophy Association of America. R. T. Moon was supported by a postdoctoral fellowship from the American Cancer Society. E. Lazarides is a recipient of a Research Career Development A ward from the National Institutes of Health.|
|Official Citation:||Canavanine inhibits vimentin assembly but not its synthesis in chicken embryo erythroid cells. R T Moon and E Lazarides 97:1309-1314. doi:10.1083/jcb.97.4.1309|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Ruth Sustaita|
|Deposited On:||17 Jul 2012 18:26|
|Last Modified:||26 Dec 2012 15:34|
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