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Procollagen mRNA metabolism during the fibroblast cell cycle and its synthesis in transformed cells
Abstract:
Procollagen mRNA was isolated from mouse embryos and used for the synthesis of a highly labelled cDNA probe complementary to collagen mRNA. This probe was used for the investigation of procollagen mRNA metabolism during the cell cycle of 3T6 mouse embryo fibroblasts in culture. Titration hybridization experiments revealed that procollagen mRNA was present throughout the cell cycle following stumulation of confluent monolayers. Procollagen mRNA levels of sparse cultures appeared similar to those of unstimulated monolayers. The fluctuating levels of collagen synthesis during the cell cycle can be ascribed to changes in the amount of collagen mRNA present. In mouse sarcoma virus transformed 3T3 cells only 20--30% of the amount of procollagen mRNA in 3T3 cells is present indicating that the decline in collagen synthesis is due to mRNA availability.
Insights
Collagen mRNA levels fluctuate during the cell cycle, impacting collagen synthesis. In transformed cells, reduced procollagen mRNA availability limits collagen production.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Collagen is a crucial structural protein synthesized by fibroblasts.
- Understanding collagen synthesis regulation is vital for tissue repair and disease research.
Purpose of the Study:
- To investigate procollagen mRNA metabolism during the fibroblast cell cycle.
- To determine the role of mRNA availability in regulating collagen synthesis.
Main Methods:
- Isolation of procollagen mRNA from mouse embryos.
- Synthesis of a labeled cDNA probe complementary to collagen mRNA.
- Titration hybridization experiments to quantify mRNA levels in cultured fibroblasts.
Main Results:
- Procollagen mRNA is present throughout the cell cycle in stimulated 3T6 fibroblasts.
- Collagen synthesis fluctuations correlate with changes in procollagen mRNA levels.
- Mouse sarcoma virus-transformed cells exhibit significantly lower procollagen mRNA levels, limiting collagen synthesis.
Conclusions:
- Collagen synthesis is regulated by the amount of available procollagen mRNA.
- Transformation by mouse sarcoma virus drastically reduces procollagen mRNA, impacting collagen production.