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Expression of growth-regulated genes in normal and SV40 transformed hamster fibroblasts
1Department of Molecular and Cell Biology, University of Kentucky, Lexington 40506-0225.
Abstract:
Transformation by the oncogenic virus SV40 has been shown to alter the expression of cellular genes at the level of RNA abundance. Many of these genes have yet to be identified. We have determined, by Northern blot analysis, the abundance levels of several growth-regulated genes in SV40-transformed cell lines to determine if their expression is altered and correlates with the ability of SV40 transformed cells to grow in low serum containing media. The mRNA abundance levels of the G1-specific genes 2A9/calcyclin, 2F1/translocase, and 4F1/vimentin were determined in the parental hamster fibroblast cell line, tk-ts13, and in two SV40 transformants, HR5 and HR8 cells, grown in medium containing 10% calf serum (normal medium) and in HR5 and HR8 cells adapted to passage in medium containing low serum. A spontaneous transformant of the parental line capable of growth in low serum in the absence of SV40 transformation (tk-ts13/1%), was also included in these studies. The low serum adapted SV40-transformed cells and the spontaneous tk-ts13 transformed cells grew more vigorously than their nonadapted counterparts in medium containing low serum. The low serum adapted cells also grew to higher saturation densities in low serum and to densities comparable to those in high serum, whereas the nonadapted cells grew to low saturation densities in low serum, but not as low as the untransformed parental.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
Simian virus 40 (SV40) transformation alters cellular gene expression, impacting cell growth in low serum conditions. SV40-transformed cells adapted to low serum exhibit enhanced proliferation and saturation density.
Area of Science:
- Molecular Biology
- Cell Biology
- Virology
Background:
- Oncogenic viruses like Simian virus 40 (SV40) can alter cellular gene expression.
- The specific cellular genes affected by SV40 transformation and their role in altered cell growth remain largely unidentified.
- Understanding these alterations is crucial for comprehending viral oncogenesis and cell transformation.
Purpose of the Study:
- To investigate the impact of SV40 transformation on the mRNA abundance of specific growth-regulated genes.
- To determine if altered gene expression in SV40-transformed cells correlates with their ability to grow in low-serum conditions.
- To compare gene expression patterns between SV40-transformed cells and a spontaneously transformed cell line capable of low-serum growth.
Main Methods:
- Northern blot analysis was used to quantify mRNA abundance levels.
- The study examined G1-specific genes: 2A9/calcyclin, 2F1/translocase, and 4F1/vimentin.
- Experiments involved a parental hamster fibroblast cell line (tk-ts13), SV40 transformants (HR5, HR8), and a spontaneous low-serum adapted transformant (tk-ts13/1%).
Main Results:
- SV40-transformed cells adapted to low serum conditions showed more vigorous growth compared to non-adapted counterparts.
- Both low-serum adapted SV40-transformed cells and the spontaneous transformant exhibited increased growth in low serum.
- Adapted cells reached higher saturation densities in low serum, comparable to high-serum conditions, unlike non-adapted cells.
Conclusions:
- SV40 transformation influences the expression of growth-regulated genes, contributing to altered cellular behavior.
- Adaptation to low-serum conditions in SV40-transformed cells involves enhanced proliferative capacity and saturation density.
- These findings highlight the complex interplay between viral transformation, gene expression, and cellular adaptation to nutrient-limited environments.