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Poly(A) polymerases in normal and virus-transformed cells.

A Pellicer, J Salas, M L Salas

    Biochimica Et Biophysica Acta
    |January 6, 1975
    PubMed
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    Researchers identified three distinct poly(A) polymerases in hamster cells, with variations observed between normal and virus-transformed cells. These enzymes exhibit specific primer and cation requirements for polyadenylation.

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Background:

    • Polyadenylation is a crucial post-transcriptional modification of eukaryotic mRNA.
    • Poly(A) polymerases (PAPs) are enzymes responsible for catalyzing polyadenylation.
    • Understanding PAPs is essential for comprehending gene expression regulation.

    Purpose of the Study:

    • To investigate the presence and characteristics of poly(A) polymerases in normal (NIL) and RNA tumor virus-transformed (NIL-HS VIRUS) hamster embryo fibroblast cells.
    • To differentiate and characterize distinct forms of poly(A) polymerases based on their properties and cellular distribution.

    Main Methods:

    • Enzyme assays were performed on cell extracts from NIL and NIL-HS VIRUS cells cultured under different conditions (dense vs. exponential growth).

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  • Chromatographic techniques were used to separate and purify different poly(A) polymerase forms.
  • Enzyme activity was characterized by varying divalent cation concentrations, primer availability, and substrate specificity.

    • Main Results:

    • Three distinct poly(A) polymerases (I, IIA, IIB) were identified in both NIL and NIL-HS VIRUS cells.
    • Forms IIA and IIB were present in both growing and dense cultures, while Poly(A) polymerase I was detected in NIL-HS VIRUS cells but not in growing NIL cells.
    • The identified poly(A) polymerases differed in chromatographic behavior and divalent cation requirements, showing high specificity for ATP and requiring a primer (cellular RNA or poly(A)).

    Conclusions:

    • The study suggests the existence of multiple poly(A) polymerase forms with distinct regulatory properties in hamster cells.
    • Viral transformation by hamster sarcoma virus influences the expression or activity of specific poly(A) polymerase forms.
    • These findings contribute to understanding the complexity of polyadenylation and its potential regulation in cellular processes and viral oncogenesis.