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Isolation of human fos-related genes and their expression during monocyte-macrophage differentiation
M Matsui1, M Tokuhara, Y Konuma
1Molecular Oncology Laboratory, Nippon Veterinary and Zootechnical College, Tokyo, Japan.
Abstract:
cDNA clones of human fos-related genes fra-1 and fra-2 were isolated by screening human c-DNA libraries with human fos DNA as a probe. We obtained human fra-1 cDNA clones that can code for a protein of 271 amino acid residues with a calculated molecular weight of 29,413 and showed 90% similarity with rat fra-1 protein. A new fos-related gene, fra-2, has one long open reading frame of 326 amino acids, and can code for a protein with a calculated molecular weight of 35,193. Two regions, a leucine zipper domain and C-terminal region, are conserved in the fos gene family. The fra-2 gene also harbors these two regions. Transcription of the fos, fra-1 and fra-2 genes was induced by phorbol ester (TPA) stimulation of U937 human monocytic cells. On TPA stimulation, the transcriptions of fos, fra-1 and fra-2 were detectable after 30, 60 and 120 min and maximum after 60, 90 and 240 min, respectively. These findings suggest that expression of the fos gene family is regulated by an orderly mechanism.
Insights
Researchers identified human fos-related genes, fra-1 and fra-2, crucial for cellular responses. Their transcription is orderly induced by TPA stimulation, suggesting a regulated mechanism in the fos gene family.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- The fos gene family plays a critical role in cellular regulation and response pathways.
- Understanding the expression patterns of fos-related genes is essential for deciphering complex biological mechanisms.
Purpose of the Study:
- To isolate and characterize human fos-related genes, specifically fra-1 and fra-2.
- To investigate the transcriptional regulation and expression kinetics of fra-1 and fra-2 in response to phorbol ester (TPA) stimulation.
Main Methods:
- Screening of human cDNA libraries using human fos DNA as a probe to isolate fra-1 and fra-2 cDNA clones.
- Bioinformatic analysis to determine protein coding potential, amino acid sequences, and molecular weights.
- Stimulation of U937 human monocytic cells with TPA to induce gene transcription and subsequent Northern blot analysis to determine expression timing.
Main Results:
- Isolation of human fra-1 and fra-2 cDNA clones, encoding proteins of 271 and 326 amino acids, respectively.
- Identification of conserved regions (leucine zipper and C-terminal) in fra-2, similar to other fos gene family members.
- TPA stimulation induced ordered transcription of fos, fra-1, and fra-2 genes, with distinct onset and peak expression times.
Conclusions:
- The human fra-1 and fra-2 genes are novel members of the fos gene family with conserved functional domains.
- Expression of the fos gene family, including fra-1 and fra-2, is regulated by a coordinated and sequential mechanism upon TPA stimulation.