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Actinomycin D binding in vitro: active chromatin preferred.

F L Yu1, W Bender

  • 1Department of Biomedical Sciences, University of Illinois College of Medicine, Rockford 61107.

Biochemistry International
|January 1, 1990
PubMed
Summary
This summary is machine-generated.

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Actinomycin D (Act. D) preferentially binds to and inhibits RNA synthesis in nucleolar chromatin, not purified DNA. This preferential effect is due to nucleolar chromosomal proteins, not the DNA itself.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Actinomycin D (Act. D) is a known inhibitor of RNA synthesis.
  • Nucleoli are key sites of ribosomal RNA synthesis within the nucleus.

Purpose of the Study:

  • To investigate the molecular basis for the preferential interaction of Actinomycin D with nucleoli.
  • To determine whether nucleolar DNA or associated proteins are responsible for Act. D's preferential binding and inhibition.

Main Methods:

  • In vitro binding assays using [3H] Actinomycin D with isolated nuclei and nucleoli.
  • Comparison of Act. D binding to purified nuclear and nucleolar DNA versus chromatin.
  • Assessment of Act. D's inhibitory effect on RNA synthesis using chromatin and purified DNA templates.

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Main Results:

  • [3H] Actinomycin D exhibited preferential binding to nucleolar chromatin compared to purified nucleolar DNA.
  • Act. D preferentially inhibited RNA synthesis in nucleolar chromatin templates.
  • These preferential effects were abolished when using purified nuclear and nucleolar DNAs.

Conclusions:

  • The preferential inhibition of nucleolar RNA synthesis by Act. D directly reflects its preferential binding to nucleolar chromatin.
  • Nucleolar chromosomal proteins, rather than nucleolar DNA, confer the preferential binding of Act. D.