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Characterization of DNA polymerase-alpha/primase complex from developing embryonic chicken brains
S Ray1, T J Kelley, L Fan
1Department of Chemistry and Biochemistry, University of Notre Dame, IN 46556.
Indian Journal of Biochemistry & Biophysics
|August 1, 1994
Summary
Researchers isolated and characterized DNA polymerase-alpha and primase activities from embryonic chicken brains. These enzymes, crucial for DNA replication initiation, were found to exist both as a complex and independently.
Area of Science:
- Molecular Biology
- Biochemistry
- Developmental Biology
Background:
- DNA polymerase-alpha and primase are essential enzymes for DNA replication.
- Understanding their interaction and regulation is key to comprehending DNA synthesis in developing tissues.
Purpose of the Study:
- To isolate, partially purify, and characterize DNA polymerase-alpha and primase activities from embryonic chicken brain.
- To investigate the association between DNA polymerase-alpha and primase.
- To explore the properties of isolated primase activity.
Main Methods:
- Enzyme isolation and partial purification using DE-23 cellulose column chromatography and immuno-affinity chromatography.
- Characterization of enzyme activities based on sensitivity to N-ethyl-maleimide, sedimentation coefficient, and isoelectric point.
- Glycerol velocity gradient centrifugation to assess enzyme complex formation.
Main Results:
- DNA polymerase-alpha and primase activities were found to exist as a complex in embryonic chicken brain extracts.
- Both complexed and free primase activities were isolated.
- Primase activity showed maximal activity with poly dC template and was not inhibited by alpha-amanitin.
Conclusions:
- The study provides insights into the complex nature of DNA polymerase-alpha and primase in embryonic chicken brain.
- Findings may contribute to understanding the regulation of chromosomal DNA replication in developing tissues.