Characterization of a candidate Trypanosoma brucei U1 small nuclear RNA gene

Appolinaire Djikeng, Ludmila Ferreira, Maximiliano D'Angelo, Pavel Dolezal, Tracey Lamb, Silvane Murta, Veronica Triggs, Sebastian Ulbert, Alejandro Villarino, Sarah Renzi, Elisabetta Ullu, Christian Tschudi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

We have previously shown that the poly(A) polymerase (PAP) gene of Trypanosoma brucei is interrupted by an intervening sequence. It was postulated that removing this intron by cis-splicing requires a yet unidentified U1 small nuclear RNA (snRNA), which in other organisms engages in base-pair interactions across the 5′ splice site during early spliceosome assembly. Here we present a characterization of a 75 nucleotide long candidate T. brucei U1 snRNA. Immunoprecipitation studies indicate that a trimethylguanosine cap structure is present at the 5′ end and that the RNA is bound to core proteins common to spliceosomal ribonucleoprotein particles. The U1 snRNA has the potential for extensive intermolecular base pairing with the PAP 5′ splice site. We used block replacement mutagenesis to identify sequences necessary for in vivo expression of U1 snRNA. We found that at least two cis-acting elements, tRNA-like A and B boxes, located in the 5′-flanking region are necessary for U1 snRNA synthesis; no internal sequences close to the transcription start site are essential, suggesting a promoter architecture distinct from other trypanosome U-snRNA genes.

Original languageEnglish
Pages (from-to)109-115
Number of pages7
JournalMolecular and Biochemical Parasitology
Volume113
Issue number1
DOIs
Publication statusPublished - 24 Mar 2001

Keywords

  • cis-splicing
  • Common core proteins
  • Intron
  • Promoter architecture
  • Trimethylguanosine cap
  • Trypanosoma brucei
  • U1 small nuclear RNA

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