Alpha-amanitin-insensitive Transcription of Variant Surface Glycoprotein Genes Provides Further Evidence for Discontinuous Transcription in Trypanosomes

Nucleic Acids Res. 1984 Dec 21;12(24):9457-72. doi: 10.1093/nar/12.24.9457.

Abstract

Many, if not all, mRNAs in T.brucei start with the same sequence of 35 nucleotides, separately encoded in clustered so-called mini-exon repeats. From these mini-exon repeats a 141-nt precursor RNA with the 35-nt sequence at its 5' end is transcribed. Indirect evidence suggests that this RNA is linked in a second step to pre-mRNA transcripts. We have studied the sensitivity of RNA synthesis to alpha-amanitin in isolated trypanosome nuclei. Transcription of several protein coding genes is almost completely inhibited by a concentration of 5 micrograms alpha-amanitin per ml, whereas strong inhibition of mini-exon transcription is achieved with 200 micrograms alpha-amanitin per ml. In contrast, transcription of genes for variant surface glycoproteins (VSGs) is not inhibited by 1000 micrograms alpha-amanitin per ml, as is transcription of the genes for the major rRNAs. Since the mature VSG mRNAs start with the 35-nt sequence, our results provide additional evidence that the 35-nt sequence and the main part of VSG mRNA are produced from independent transcription units, these are transcribed by (partly) different RNA polymerases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amanitins / pharmacology*
  • Animals
  • Cell Nucleus / physiology
  • DNA-Directed RNA Polymerases / antagonists & inhibitors
  • Glycoproteins / genetics*
  • RNA Polymerase II / metabolism
  • RNA Polymerase III / metabolism
  • Transcription, Genetic* / drug effects
  • Trypanosoma brucei brucei / genetics*
  • Variant Surface Glycoproteins, Trypanosoma

Substances

  • Amanitins
  • Glycoproteins
  • Variant Surface Glycoproteins, Trypanosoma
  • RNA Polymerase II
  • DNA-Directed RNA Polymerases
  • RNA Polymerase III