Cloning of murine CDK9/PITALRE and its tissue-specific expression in development
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L Bagella1, T K MacLachlan, R J Buono
1Department of Pathology, Anatomy and Cell Biology, Sbarro Institute for Cancer Research and Molecular Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Abstract:
The cdc2-family of serine/threonine kinases and their binding partners recently were implicated in developmental roles. We previously cloned a cdc2-related kinase, cdk9/PITALRE, that is able to phosphorylate the retinoblastoma protein in vitro. We describe here the cloning and the characterization of the mouse homolog of cdk9/PITALRE. The murine cDNA is 98% identical with humans and is expressed at high levels in brain and kidney tissues. The kinase activity and protein expression of cdk9/PITALRE were highest in terminally differentiated tissues such as the muscle and brain. In situ immunohistology and immunofluorescence detected cdk9/PITALRE protein not only within terminally differentiated cells such as muscle and neuronal cells, but also in proliferating cells. C2C12 and P19 cells induced to differentiate along muscle and neural lineages peaked in cdk9/PITALRE kinase activity at the end of differentiation. These results suggest that, among other roles, cdk9/PITALRE plays a role not unlike cdk5 in the differentiation of certain cell types.
Insights
The study identifies the mouse cdk9/PITALRE kinase, crucial for cell differentiation. Its expression and activity peak in terminally differentiated tissues, suggesting a role similar to cdk5 in development.
Area of Science:
- Molecular Biology
- Cell Biology
- Developmental Biology
Background:
- The cdc2-family of kinases plays significant roles in cellular development.
- cdk9/PITALRE, a cdc2-related kinase, is known to phosphorylate the retinoblastoma protein.
- Previous cloning of human cdk9/PITALRE established its potential involvement in cellular processes.
Purpose of the Study:
- To clone and characterize the mouse homolog of cdk9/PITALRE.
- To investigate the expression patterns and kinase activity of mouse cdk9/PITALRE during differentiation.
- To elucidate the potential role of cdk9/PITALRE in cell differentiation, particularly in muscle and neural lineages.
Main Methods:
- Cloning of the murine cDNA for cdk9/PITALRE.
- Analysis of cDNA sequence identity with human counterpart.
- Quantitative assessment of gene expression in various mouse tissues.
- In situ immunohistology and immunofluorescence to detect protein localization.
- Measurement of kinase activity in differentiating cell lines (C2C12 and P19).
Main Results:
- Murine cdk9/PITALRE cDNA shows 98% identity to human cDNA.
- High expression levels of cdk9/PITALRE were observed in brain and kidney tissues.
- Kinase activity and protein expression were highest in terminally differentiated tissues like muscle and brain.
- cdk9/PITALRE protein was detected in both terminally differentiated and proliferating cells.
- Peak kinase activity in C2C12 and P19 cells correlated with the end of muscle and neural differentiation.
Conclusions:
- Mouse cdk9/PITALRE is highly conserved with its human homolog.
- cdk9/PITALRE expression and activity are linked to terminal differentiation.
- The findings suggest cdk9/PITALRE participates in the differentiation of specific cell types, similar to cdk5.
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