Cloning, chromosomal localization and expression analysis of the mouse Akt2 oncogene
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D A Altomare1, K Guo, J Q Cheng
1Department of Medical Oncology, Fox Chase Cancer Center, Philadelphia, Pennsylvania 19111, USA.
Abstract:
We isolated mouse cDNA clones containing the entire coding region of the putative oncogene Akt2. Sequence analysis revealed that, like its human homolog, Akt2 encodes a protein-serine/threonine kinase containing a pleckstrin homology domain at its amino terminus. Fluorescence in situ hybridization of the mouse cDNA to rodent metaphase spreads demonstrated that the Akt2 gene maps to mouse chromosome band 7B1 and rat chromosome 1q22. Expression levels of mouse Akt2 mRNA and Akt2 protein varied among tissues, with the highest levels in skeletal muscle. Akt2 expression was low in a multipotent fibroblast cell line, but it was upregulated when these cells were transformed with Myod and induced to differentiate into myocytes. These data demonstrate that Akt2 expression is activated during cellular differentiation and suggest that it functions in the signaling pathways of some adult tissues.
Insights
The Akt2 gene, a protein-serine/threonine kinase, is mapped to mouse chromosome 7B1. Akt2 expression is highest in skeletal muscle and activated during cellular differentiation.
Area of Science:
- Molecular Biology
- Genetics
- Cellular Biology
Background:
- The Akt2 gene encodes a protein-serine/threonine kinase with a pleckstrin homology domain, similar to its human homolog.
- Understanding the Akt2 gene's function and regulation is crucial for insights into cellular signaling and tissue-specific roles.
Purpose of the Study:
- To isolate and characterize mouse cDNA clones of the Akt2 gene.
- To determine the chromosomal mapping of the Akt2 gene in rodents.
- To investigate the expression patterns of Akt2 mRNA and protein in various tissues and during cellular differentiation.
Main Methods:
- Isolation of mouse cDNA clones encoding the full coding region of Akt2.
- Sequence analysis to identify protein domains and homology to human Akt2.
- Fluorescence in situ hybridization (FISH) for chromosomal mapping.
- Quantitative analysis of Akt2 mRNA and protein expression levels in different tissues and cell lines.
- Induction of differentiation in a multipotent fibroblast cell line using Myod.
Main Results:
- Mouse Akt2 cDNA clones containing the entire coding region were successfully isolated.
- Sequence analysis confirmed Akt2 encodes a protein-serine/threonine kinase with a pleckstrin homology domain.
- FISH mapped the Akt2 gene to mouse chromosome band 7B1 and rat chromosome 1q22.
- Akt2 mRNA and protein expression varied by tissue, with highest levels observed in skeletal muscle.
- Akt2 expression was low in multipotent fibroblasts but significantly upregulated upon Myod-induced differentiation into myocytes.
Conclusions:
- The Akt2 gene is located on mouse chromosome 7B1 and rat chromosome 1q22.
- Akt2 exhibits tissue-specific expression, notably high in skeletal muscle.
- Akt2 expression is activated during cellular differentiation, suggesting a role in this process.
- The findings indicate Akt2 functions within signaling pathways relevant to adult tissues and cellular differentiation.