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Expression of candidate cell death genes in cell lines during apoptosis
1Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Victoria, Australia.
Abstract:
We have cloned mouse candidate cell death genes RP-2 and nedd2 and used Northern blot analysis to study their expression in a growth-factor-dependent cell line (FDC-P1) that can be induced to undergo apoptosis by growth factor withdrawal and in a thymoma line (ST4) that undergoes apoptosis when irradiated. RP-2 was expressed in FDC-P1 cells even when not undergoing apoptosis, and mRNA levels did not increase when apoptosis was induced by growth factor withdrawal. FDC-P1 cells expressed two genes that are similar to the Caenorhabditis elegans cell death gene ced-3: a 3.7 kilobase (kb) nedd2 message and low levels of a 5-kb message for interleukin-1 beta converting enzyme. Levels of these messages did not change with the induction of cell death, but transfection of fibroblasts with constructs expressing nedd2 caused them to undergo apoptosis. These results suggest that expression of RP-2 and nedd2 is not sufficient for apoptosis, and if the products of these genes are involved in apoptosis of FDC-P1 cells, they are regulated by posttranslational mechanisms.
Insights
Researchers investigated cell death genes RP-2 and nedd2. Their expression did not directly correlate with apoptosis induction, suggesting post-translational regulation is key for programmed cell death.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Apoptosis, or programmed cell death, is crucial for development and tissue homeostasis.
- Understanding the genetic regulation of apoptosis is vital for disease research.
Purpose of the Study:
- To investigate the expression and role of candidate cell death genes RP-2 and nedd2 in apoptosis.
- To determine if RP-2 and nedd2 expression levels correlate with apoptosis induction in specific cell lines.
Main Methods:
- Cloning of mouse candidate cell death genes RP-2 and nedd2.
- Northern blot analysis to study gene expression in FDC-P1 and ST4 cell lines.
- Transfection of fibroblasts with nedd2 constructs.
Main Results:
- RP-2 mRNA levels were consistent in FDC-P1 cells, regardless of apoptosis induction.
- Nedd2 and interleukin-1 beta converting enzyme (ICE) mRNA levels did not change during apoptosis induction.
- Transfection with nedd2 induced apoptosis in fibroblasts, but RP-2 and nedd2 expression alone were insufficient for apoptosis in FDC-P1 cells.
Conclusions:
- RP-2 and nedd2 gene expression is not sufficient to trigger apoptosis.
- The products of RP-2 and nedd2 may be involved in apoptosis, but are likely regulated by post-translational mechanisms.