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Decrease in ADP-ribosylation of HeLa non-histone proteins from interphase to metaphase.

K W Adolph, M K Song

    Biochemistry
    |January 15, 1985
    PubMed
    Summary
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    ADP-ribosylation of non-histone proteins differs significantly between interphase and metaphase cells. During mitosis, poly(ADP-ribose) polymerase becomes the primary target, suggesting a structural role in chromosome organization.

    Area of Science:

    • Cell Biology
    • Biochemistry
    • Molecular Biology

    Background:

    • ADP-ribosylation is a post-translational modification crucial for various cellular processes.
    • Non-histone proteins play diverse roles in nuclear structure and function.
    • Significant changes in protein modification occur during the cell cycle, particularly mitosis.

    Purpose of the Study:

    • To investigate variations in ADP-ribosylation patterns of non-histone proteins between interphase and metaphase cells.
    • To identify major ADP-ribosylated non-histone proteins in different cellular states.
    • To explore the potential structural implications of ADP-ribosylation during mitosis.

    Main Methods:

    • Utilized 32P-labeled nicotinamide adenine dinucleotide ([32P]NAD) as a specific precursor for radioactive labeling.

    Related Experiment Videos

  • Incubated permeabilized interphase and mitotic cells, isolated nuclei, and chromosomes with the label.
  • Analyzed labeled proteins using one-dimensional and two-dimensional gel electrophoresis.
  • Employed ADP-ribosylation inhibitors and snake venom phosphodiesterase to validate modification and removal.

    • Main Results:

    • Over 100 non-histone protein species were ADP-ribosylated in interphase nuclei and chromatin.
    • Major ADP-ribosylated proteins in interphase included lamins and poly(ADP-ribose) polymerase.
    • Metaphase chromosomes exhibited a fundamentally different pattern, with poly(ADP-ribose) polymerase as the sole major acceptor.
    • Labeling conditions affected incorporation levels and distribution but yielded generally similar patterns in interphase.

    Conclusions:

    • ADP-ribosylation of non-histone proteins undergoes a profound redistribution during mitosis.
    • The shift in ADP-ribosylation targets suggests a significant structural role for this modification in metaphase chromosomes.
    • Further research is warranted to elucidate the precise structural functions of ADP-ribosylated non-histone proteins during cell division.