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Cloning of the rat proto-oncogene bmi-1
1Department of Pathology, Faculty of Medicine, Kyoto University, Japan.
Abstract:
The bmi-1 gene was identified as a common proviral integration site in Moloney murine leukemia virus. In the present studies, we cloned and sequenced the rat bmi-1 gene by reverse transcriptase-polymerase chain reaction (RT-PCR) using degenerate PCR primers of homologous sequences between mouse and human. We found 93% identity to the mouse bmi-1 cDNA and 90% identity to the human bmi-1. The open reading frame encodes a protein of 324 amino acids. In the deduced amino acid sequence we observed 95% and 94% homology to the mouse and human, respectively. The structural motifs, a novel zinc finger motif and a putative helix-turn-helix motif, were conserved in the predicted rat BMI-1 protein. We also confirmed ubiquitous expression of bmi-1 in normal tissues except brain. These results suggest functional conservation of the bmi-1 gene in the rat.
Insights
Researchers cloned and sequenced the rat bmi-1 gene, finding high similarity to mouse and human sequences. This suggests the bmi-1 gene is functionally conserved across these species.
Area of Science:
- Molecular Biology
- Genetics
- Comparative Genomics
Background:
- The bmi-1 gene is a known integration site for Moloney murine leukemia virus.
- Understanding bmi-1's role is crucial for cancer research and viral oncogenesis.
Purpose of the Study:
- To clone and sequence the rat bmi-1 gene.
- To analyze the structural and functional conservation of the rat bmi-1 gene compared to its mouse and human counterparts.
Main Methods:
- Reverse transcriptase-polymerase chain reaction (RT-PCR) was employed using degenerate primers.
- Sequence analysis was performed to determine cDNA and amino acid homology.
- Expression patterns were analyzed in normal rat tissues.
Main Results:
- The rat bmi-1 cDNA showed 93% identity to mouse and 90% identity to human cDNA.
- The deduced rat BMI-1 protein (324 amino acids) exhibited 95% homology to mouse and 94% homology to human proteins.
- Conserved structural motifs, including a zinc finger and a helix-turn-helix motif, were identified.
- Ubiquitous expression of bmi-1 was observed in normal tissues, excluding the brain.
Conclusions:
- The rat bmi-1 gene shares significant sequence and structural homology with mouse and human bmi-1.
- These findings strongly suggest functional conservation of the bmi-1 gene in rats.
- This conservation implies similar roles in cellular processes and potential implications for oncogenesis studies.