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Isolation of retinoic acid-repressed genes from P19 embryonal carcinoma cells
H Nakshatri1, P Bouillet, P Bhat-Nakshatri
1Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, Collège de France, Strasbourg, France.
Abstract:
Retinoic acid (RA) plays a critical role in normal development, growth and differentiation by modulating the expression of target genes. Using substractive hybridization cloning, we isolated two cDNAs, whose corresponding mRNAs are repressed upon RA treatment of P19 embryonal carcinoma (EC) cells. The cDNAs correspond to the serine hydroxymethyltransferase (shmt) gene and the early transposon, ETnMG1. RA appears to reduce the stability of ETnMG1 transcript. We also report the sequence of two different isoforms of mouse SHMT. Since SHMT activity is increased when cells are stimulated to proliferate and during the S phase of the cell cycle, we suggest that repression of shmt expression is an important step in RA-induced cell growth arrest and differentiation.
Insights
Retinoic acid (RA) represses serine hydroxymethyltransferase (SHMT) gene expression in P19 embryonal carcinoma cells. This repression is linked to RA-induced cell growth arrest and differentiation, impacting cell proliferation.
Area of Science:
- Developmental Biology
- Molecular Biology
- Gene Regulation
Background:
- Retinoic acid (RA) is crucial for normal development, growth, and differentiation.
- RA modulates the expression of target genes, influencing cellular processes.
Purpose of the Study:
- To identify genes repressed by RA in P19 embryonal carcinoma (EC) cells.
- To investigate the role of serine hydroxymethyltransferase (SHMT) in RA-induced cellular responses.
Main Methods:
- Subtractive hybridization cloning was used to isolate differentially expressed cDNAs.
- mRNA stability was assessed following RA treatment.
- Sequencing identified two isoforms of mouse SHMT.
Main Results:
- Two cDNAs, corresponding to the SHMT gene and the ETnMG1 transposon, were repressed by RA.
- RA treatment reduced the stability of the ETnMG1 transcript.
- Two distinct mouse SHMT isoforms were sequenced.
Conclusions:
- Repression of SHMT gene expression is a key event in RA-induced cell growth arrest.
- The findings suggest a role for SHMT in RA-mediated differentiation processes.
- Modulation of SHMT activity may be a mechanism by which RA controls cell proliferation and differentiation.