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Cyclin D2 interacts with cdk-5 and modulates cellular cdk-5/p35 activity
S Guidato1, D M McLoughlin, A J Grierson
1Department of Clinical Neurosciences, Institute of Psychiatry, London, England.
Abstract:
Cyclin-dependent kinase-5 (cdk-5) is a serine/threonine kinase that displays neurone-specific activity. Experimental manipulation of cdk-5 expression in neurones has shown that cdk-5 is essential for proper development of the nervous system and, in particular, for outgrowth of neurites. Such observations suggest that cdk-5 activity must be tightly controlled during development of the nervous system. To identify possible regulators of cdk-5, we used the yeast two-hybrid system to search for proteins that interact with cdk-5. In two independent yeast transformation events, cyclin D2 interacted with cdk-5. Immunoprecipitation experiments confirmed that cyclin D2 and cdk-5 interact in mammalian cells. Cyclin D2 did not activate cdk-5 as assayed using three different substrates, which was in contrast to a known cdk-5 activator, p35. However, cyclin D2 expression led to a decrease in cdk-5/p35 activity in transfected cells. As cyclin D2 and cdk-5 are known to share overlapping patterns of expression during development of the CNS, the results presented here suggest a role for cyclin D2 in modulating cdk-5 activity in postmitotic developing neurones.
Insights
Cyclin-dependent kinase-5 (CDK-5) is crucial for nervous system development. Researchers found that cyclin D2 interacts with CDK-5 and may regulate its activity in developing neurons.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Cyclin-dependent kinase-5 (CDK-5) is a neuron-specific kinase vital for nervous system development and neurite outgrowth.
- Tight regulation of CDK-5 activity is essential during neural development.
Purpose of the Study:
- To identify novel regulators of CDK-5 activity.
- To investigate the interaction between CDK-5 and cyclin D2.
Main Methods:
- Yeast two-hybrid system to screen for CDK-5 interacting proteins.
- Immunoprecipitation experiments in mammalian cells.
- Enzyme activity assays using different substrates.
Main Results:
- Cyclin D2 was identified as a CDK-5 interacting protein through yeast two-hybrid screening.
- Interaction between cyclin D2 and CDK-5 was confirmed in mammalian cells.
- Cyclin D2 did not activate CDK-5 but decreased CDK-5/p35 activity in transfected cells.
Conclusions:
- Cyclin D2 interacts with CDK-5 in mammalian cells.
- Cyclin D2 may play a role in modulating CDK-5 activity in developing neurons.
- Findings suggest a novel regulatory mechanism for CDK-5 during central nervous system development.