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Related Experiment Videos

RNA polymerase I transcription factor Rrn3 is functionally conserved between yeast and human.

B Moorefield1, E A Greene, R H Reeder

  • 1Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, WA 98109, USA.

Proceedings of the National Academy of Sciences of the United States of America
|April 12, 2000
PubMed
Summary
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Researchers cloned a human RNA polymerase I transcription factor, Rrn3, functionally conserved in yeast and mammals. This discovery suggests a shared mechanism for regulating ribosomal RNA synthesis across eukaryotes.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • RNA polymerase I (Pol I) is crucial for synthesizing ribosomal RNA (rRNA).
  • Transcription factors regulate Pol I activity, but their conservation across species is not fully understood.
  • Rrn3 is a known Pol I transcription factor in yeast.

Purpose of the Study:

  • To identify and characterize a human homolog of the yeast Rrn3 transcription factor.
  • To investigate the functional conservation of Rrn3 between yeast and mammals.
  • To explore the implications for rRNA synthesis regulation.

Main Methods:

  • Cloning of a human cDNA encoding a putative Rrn3 homolog.
  • Expression and purification of the recombinant human protein.

Related Experiment Videos

  • Functional assays in a yeast strain with a disrupted RRN3 gene (in vivo rescue).
  • Site-directed mutagenesis of conserved amino acids.

    • Main Results:

    • A human cDNA related to yeast Rrn3 was successfully cloned.
    • The recombinant human Rrn3 protein showed sequence similarity and immunological cross-reactivity to yeast Rrn3.
    • The human Rrn3 functionally rescued the yeast RRN3 disruption mutant.
    • A conserved point mutation abolished the function of both yeast and human Rrn3, confirming functional significance.

    Conclusions:

    • Human Rrn3 is functionally conserved with yeast Rrn3.
    • This marks the first RNA polymerase I-specific transcription factor found to be conserved between yeast and mammals.
    • Suggests conserved mechanisms for regulating ribosomal RNA synthesis in eukaryotes.