Chromatin immunoprecipitation and microarray analysis reveal that TFIIB occupies the SL RNA gene promoter region in Trypanosoma brucei chromosomes

Wenzhe Liu, Anish Das, Rachel Morales, Mahrukh Banday, Virginie Aris, David Lukac, Patricia Soteropoulos, David A. Wah, Jennifer Palenchar, Vivian Bellofatto

Research output: Contribution to journalArticle

Abstract

RNA polymerase II (RNAP-II) synthesizes the m7G-capped Spliced Leader (SL) RNA and most protein-coding mRNAs in trypanosomes. RNAP-II recruitment to DNA usually requires a set of transcription factors that make sequence-specific contacts near transcriptional start sites within chromosomes. In trypanosomes, the transcription factor TFIIB is necessary for RNAP-II-dependent SL RNA transcription. However, the trypanosomal TFIIB (tTFIIB) lacks the highly basic DNA binding region normally found in the C-terminal region of TFIIB proteins. To assess the precise pattern of tTFIIB binding within the SL RNA gene locus, as well as within several other loci, we performed chromatin immunoprecipitation/microarray analysis using a tiled gene array with a probe spacing of 10 nucleotides. We found that tTFIIB binds non-randomly within the SL RNA gene locus mainly within a 220-nt long region that straddles the transcription start site. tTFIIB does not bind within the small subunit (SSU) rRNA locus, indicating that trypanosomal TFIIB is not a component of an RNAP-I transcriptional complex. Interestingly, discrete binding sites were observed within the putative promoter regions of two loci on different chromosomes. These data suggest that although trypanosomal TFIIB lacks a highly basic DNA binding region, it nevertheless localizes to discrete regions of chromatin that include the SL RNA gene promoter.

Original languageEnglish (US)
Pages (from-to)139-142
Number of pages4
JournalMolecular and Biochemical Parasitology
Volume186
Issue number2
DOIs
StatePublished - Dec 1 2012

All Science Journal Classification (ASJC) codes

  • Parasitology
  • Molecular Biology

Keywords

  • Chromatin immunoprecipitation
  • RNA synthesis
  • SL RNA genes
  • TFIIB
  • Trypanosoma brucei
  • Trypanosomes

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