Production of GP140, a major component of the extracellular matrix of cultured fibroblasts, is markedly decreased in SV40 transformed cells as compared with normal cells (Carter, W. G., 1982, J. Biol. Chem., 257:13805-13815). To determine at what step the biosynthesis is inhibited, we compared the levels of functional mRNA for GP140 in normal and transformed fibroblasts. Translation of total RNA from W138 cells in a reticulocyte lysate, followed by immunoprecipitation with affinity-purified antibodies to GP140, yielded a single polypeptide with an Mr of 125,000. This polypeptide was identified as GP140 based on its immunoreactivity, collagenase sensitivity, and comigration on polyacrylamide gels with GP140 synthesized by cells in the presence of tunicamycin and 2,2'-bipyridyl. No cell-free synthesis of GP140 was observed with total RNA from SV40 transformed W138 cells, indicating that these cells contain very low levels of GP140-specific mRNA. The biosynthesis of GP140 might therefore be blocked at the transcriptional level.
Article| February 01 1985
Translatable mRNA for GP140 (a subunit of type VI collagen) is absent in SV40 transformed fibroblasts.
J B Lewis
W G Carter
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1985) 100 (2): 638–641.
B Trüeb, J B Lewis, W G Carter; Translatable mRNA for GP140 (a subunit of type VI collagen) is absent in SV40 transformed fibroblasts.. J Cell Biol 1 February 1985; 100 (2): 638–641. doi: https://doi.org/10.1083/jcb.100.2.638
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