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Nucleotide polymerases in the developing avian erythrocyte

S S Longacre, W J Rutter

    The Journal of Biological Chemistry
    |January 10, 1977
    PubMed
    Summary

    This study characterizes nucleotide polymerase activities in developing avian erythroid cells, revealing declining patterns that correlate with cellular maturation and suggesting polymerase levels control gene expression and DNA replication during development.

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    Area of Science:

    • Molecular Biology
    • Cell Biology
    • Biochemistry

    Background:

    • Avian erythroid cells offer a model system for studying cellular development.
    • Understanding the enzymes involved in nucleic acid synthesis is crucial for comprehending cellular processes.

    Purpose of the Study:

    • To investigate the activities and characteristics of DNA-dependent RNA polymerases, RNA-dependent terminal ribonucleotidyltransferases, and DNA-dependent DNA polymerases during avian erythroid cell development.
    • To correlate the activity levels of these nucleotide polymerases with the maturation stages of red blood cells.

    Main Methods:

    • Separation of avian erythroid cells into five developmental stages using albumin density gradients.
    • Partial purification and characterization of solubilized enzyme activities via ion exchange chromatography, ion filtration chromatography, and velocity sedimentation.
    • Analysis of three types of nucleotide polymerases: DNA-dependent RNA polymerases (I, II, III), RNA-dependent terminal ribonucleotidyltransferases, and DNA-dependent DNA polymerases (alpha, beta, and a third distinct entity).

    Main Results:

    • Identified DNA-dependent RNA polymerases I and II, with minimal Polymerase III activity.
    • Detected at least two species of RNA-dependent terminal ribonucleotidyltransferases, including poly(A) polymerase and another requiring an RNA primer.
    • Resolved three DNA polymerase activities, including alpha, beta, and a novel third polymerase, with poly(dC)-dependent RNA polymerase activity associated with the alpha polymerase enriched in the third DNA polymerase species.
    • Observed characteristic declining activity patterns for all investigated nucleotide polymerases as erythroid cells matured.

    Conclusions:

    • The observed decline in nucleotide polymerase activity during avian erythroid cell maturation aligns with known synthetic rates of their in vivo products.
    • Changes in the levels of these polymerases likely play a significant role in regulating transcription and replication processes during cellular development.

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