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Identification of an activator required for elevation of maturation-promoting factor (MPF) activity by gamma-S-ATP
1Department of Pharmacology, University of Colorado School of Medicine, Denver 80262.
Abstract:
Maturation-promoting factor (MPF) is a cell cycle control element able to cause cells to enter M-phase upon microinjection and will induce metaphase in nuclei incubated in cell extracts. Previous work has shown that MPF is composed of a complex between p34cdc 2 protein kinase and a B-type cyclin. In the present work gamma-S-ATP was found to cause activation of MPF activity in partially purified preparations, but this activation was lost upon chromatography on Matrex Green gel A. Readdition of other Matrex Green fractions to purified MPF restored the ability of gamma-S-ATP to activate MPF for nuclear breakdown as well as phosphorylation of histone H1. Use of the system described here will facilitate study of p34cdc 2 kinase activation and identification of elements involved in MPF regulation.
Insights
Maturation-promoting factor (MPF) is crucial for cell cycle control. This study identified key components that restore MPF activity, aiding research into cell cycle regulation and p34cdc2 kinase activation.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Maturation-promoting factor (MPF) regulates entry into M-phase of the cell cycle.
- MPF comprises a complex of p34cdc2 protein kinase and a B-type cyclin.
Purpose of the Study:
- To investigate the activation of MPF using gamma-S-ATP.
- To identify regulatory elements involved in MPF activation and p34cdc2 kinase activity.
Main Methods:
- Partial purification of MPF.
- Chromatography on Matrex Green gel A.
- Assays for MPF activity, nuclear breakdown, and histone H1 phosphorylation.
Main Results:
- Gamma-S-ATP activated MPF in partially purified preparations.
- MPF activation by gamma-S-ATP was lost after chromatography.
- Readdition of specific Matrex Green fractions restored gamma-S-ATP-dependent MPF activation.
Conclusions:
- Specific fractions are required for gamma-S-ATP-mediated MPF activation.
- This system facilitates the study of p34cdc2 kinase activation and MPF regulation.