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Published on: August 19, 2014
Identification of a novel mouse p53 target gene DDA3
1Institute of Biochemistry, National Yang Ming University, Shih-Pai, Taipei, Taiwan 112, Republic of China.
Abstract:
We have identified a novel p53 regulated gene designated DDA3 through differential mRNA display on IW32 erythroleukemia cells containing a temperature sensitive p53 allele, tsp53val-135. DDA3 mRNA induction could be observed in all sublines expressing tsp53val-135 cultured at permissive temperature as well as in NIH3T3 cells undergoing DNA damage. Upregulation of DDA3 could be detected within 2 h after down-shifting the temperature to 32.5 degrees C; upon shifting back to 38.5 degrees C, DDA3 mRNA rapidly degraded with a half-life of less than 2 h. Actinomycin D, but not cycloheximide, inhibited the p53 dependent DDA3 induction, suggesting that the activation is through transcriptional regulation and does not require de novo protein synthesis. DDA3 was expressed in multiple mouse tissues including brain, spleen, lung, kidney and testis. Full-length DDA3 cDNA was cloned and it contained an open reading frame predicted to encode a proline rich protein of 329 amino acids. Overexpression of DDA3 in H1299 lung carcinoma cells suppressed colony formation. These results suggest that DDA3 is a p53-regulated gene that might participate in the p53-mediated growth suppression.
Insights
Researchers discovered DDA3, a novel gene regulated by p53. This gene
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Regulation
Background:
- The p53 tumor suppressor protein plays a critical role in cellular responses to stress.
- Identifying p53 target genes is crucial for understanding its tumor-suppressive functions.
Purpose of the Study:
- To identify novel genes regulated by p53.
- To characterize the function of a newly identified p53-regulated gene, DDA3.
Main Methods:
- Differential mRNA display using erythroleukemia cells with a temperature-sensitive p53 allele.
- Analysis of DDA3 mRNA induction and degradation kinetics.
- Investigating the mechanism of DDA3 regulation (transcriptional vs. protein synthesis).
- Cloning of full-length DDA3 cDNA and functional studies via overexpression.
Main Results:
- A novel p53-regulated gene, DDA3, was identified.
- DDA3 mRNA is rapidly induced by p53 activation and degraded upon p53 inactivation.
- DDA3 induction is transcriptionally regulated and does not require new protein synthesis.
- DDA3 is expressed in various mouse tissues and its overexpression suppresses cancer cell growth.
Conclusions:
- DDA3 is a novel p53 target gene.
- DDA3 may play a role in p53-mediated growth suppression and tumor suppression.
