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Levels of RNA polymerase activities during growth, encystment and germination ofPhysarum polycephalum.

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Sequence complexity of poly(A) RNA fromPhysarum polycephalum.

Annette Baeckmann1, Helmut W Sauer1

  • 1Zoologisches Institut der Universität, Röntgenring 10, D-8700, Würzburg, Germany.

Wilhelm Roux'S Archives of Developmental Biology
|March 18, 2017
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Summary

Physarum poly(A) RNA analysis reveals similar polypeptide patterns across cell cycle phases and starvation. Gene expression changes significantly, with substantial RNA sequence reduction during starvation.

Keywords:
DevelopmentPhysarumRNASequence complexity

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Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Polyadenylated (poly(A)) RNA plays a crucial role in gene expression regulation.
  • Understanding mRNA sequence dynamics during different cellular states is vital for comprehending cellular processes.

Purpose of the Study:

  • To investigate the complexity and changes in mRNA populations during the cell cycle (S and G2 phases) and starvation in Physarum macroplasmodia.
  • To quantify the proportion of the genome transcribed and the abundance of specific RNA species under different conditions.

Main Methods:

  • In vitro translation and fluorography to analyze polypeptide patterns from poly(A) RNA.
  • DNA reassociation kinetics (Cot) for isolating single-copy DNA.
  • Saturation hybridization of labeled DNA with poly(A) RNA.
  • Homologous and heterologous cDNA/poly(A) RNA hybridization reactions (Rot) to assess transcriptional complexity and sequence overlap.

Main Results:

  • Similar polypeptide patterns were observed from poly(A) RNA of S, G2, and starved Physarum macroplasmodia.
  • 23% of the genome is transcribed during growth and 17% during starvation, based on DNA/poly(A) RNA hybridization.
  • 22-28% of the genome is transcribed during growth and 12% during starvation, with 50-80 RNA species present at ~1,000 copies/nucleus.
  • Up to 25,000 RNA species (1-5 copies/nucleus) during growth and ~15,000 during starvation were estimated.
  • RNA sequences in S and G2 phases are similar, with higher abundance in G2. About 25% of growth-related RNA sequences are undetectable during starvation.

Conclusions:

  • Physarum exhibits significant modulation of gene expression between growth and starvation states.
  • While overall polypeptide patterns remain similar, substantial changes occur in the abundance and diversity of mRNA sequences.
  • Cell cycle progression involves dynamic regulation of RNA populations, with distinct differences observed between growth and starvation conditions.