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Related Experiment Videos

Alu transcripts: cytoplasmic localisation and regulation by DNA methylation

W M Liu1, R J Maraia, C M Rubin

  • 1Section of Molecular and Cellular Biology, University of California, Davis 95616.

Nucleic Acids Research
|March 25, 1994
PubMed
Summary

HeLa cells transcribe full-length Alu elements using RNA polymerase III. Hypomethylation of Alu DNA increases these transcript levels, revealing insights into gene regulation.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Epigenetics

Background:

  • Alu elements are abundant repetitive sequences in the human genome.
  • Transcription of Alu elements is typically repressed.
  • RNA polymerase III (Pol III) transcribes various small RNAs.

Purpose of the Study:

  • To investigate the transcription of full-length Alu elements in HeLa cells.
  • To characterize the nature and regulation of these Alu transcripts.
  • To understand the role of DNA methylation in Alu transcription.

Main Methods:

  • Primer extension using reverse transcriptase to detect Alu transcripts.
  • Analysis of cloned complementary DNA (cDNA) sequences.
  • Sucrose gradient sedimentation to assay for ribonucleoprotein particles (RNPs).

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  • Treatment with 5-azacytidine to induce DNA hypomethylation.

    • Main Results:

    • Full-length Alu transcripts are detected and originate from internal Pol III promoters.
    • These transcripts are found in cytoplasmic polyadenylated RNAs and form RNPs.
    • Alu transcript abundance increases following 5-azacytidine treatment, correlating with DNA hypomethylation.
    • Sequence analysis reveals transcription of diverse Alu repeats and conservation of the Pol III promoter A box in some transcripts.

    Conclusions:

    • HeLa cells actively transcribe full-length Alu elements via Pol III.
    • DNA hypomethylation, induced by 5-azacytidine, enhances Alu transcript levels.
    • The findings shed light on the regulation of repetitive element transcription and its epigenetic control.