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Related Experiment Videos

Nucleic acid polymerizing enzymes in developing Strongylocentrotus franciscanus embryos

P W Morris, W J Rutter

    Biochemistry
    |July 13, 1976
    PubMed
    Summary

    Sea urchin eggs contain essential DNA polymerase, RNA polymerase, and terminal riboadenylate transferase (TRT) enzymes for development. Post-fertilization increases in DNA and RNA synthesis result from activating these pre-existing egg enzymes.

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    Area of Science:

    • Molecular Biology
    • Developmental Biology
    • Biochemistry

    Background:

    • Fertilization triggers rapid increases in DNA, RNA, and polyadenylated RNA synthesis in sea urchin embryos.
    • Understanding the enzymatic machinery responsible for these increases is crucial for comprehending early development.

    Purpose of the Study:

    • To investigate the presence and localization of DNA polymerase, RNA polymerase, and terminal riboadenylate transferase (TRT) activities in sea urchin (Strongylocentrotus franciscanus) eggs and embryos.
    • To determine if enzyme synthesis or activation accounts for increased nucleic acid synthesis post-fertilization.

    Main Methods:

    • DEAE-Sephadex chromatography was used to analyze whole cell extracts from sea urchin eggs and staged embryos.
    • Enzyme activities, including DNA-dependent RNA polymerase, DNA-dependent DNA polymerase, and TRT, were measured.

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  • Sensitivity to alpha-amanitin and gradient sievorptive elution were employed to characterize RNA polymerase forms (I, II, and III).
  • Nucleate and anucleate egg halves were used to assess enzyme localization.

    • Main Results:

    • All three enzyme activities (DNA polymerase, RNA polymerase, TRT) are present in unfertilized eggs and remain constant in total activity per embryo throughout development.
    • Increased nucleic acid synthesis post-fertilization is attributed to the activation of pre-existing egg enzymes, not de novo synthesis.
    • RNA polymerases are initially distributed in both the cytoplasm and nucleus, becoming progressively nuclear during development.
    • Early embryos have low RNA polymerase II activity, while later stages show more balanced activities of RNA polymerases I, II, and III.
    • Two distinct RNA polymerase III species were detected, with one appearing only in later developmental stages.

    Conclusions:

    • Sea urchin eggs possess sufficient polymerase and TRT activities for embryogenesis, with post-fertilization increases driven by enzyme activation.
    • Dynamic changes in RNA polymerase composition, including the appearance of new RNA polymerase III species, occur during sea urchin development.
    • These findings suggest enzyme interconversion through subunit modification or differential synthesis/degradation contributes to developmental regulation.