Abstract

Trypanosomes are ancient eukaryotic parasites in which the protein-coding genes, organized in large polycistronic clusters on both strands, are transcribed from as yet unidentified promoters. In an effort to reveal transcriptional initiation sites, we examined the Trypanosoma cruzi genome for histone modification patterns shown to be linked to active genes in various organisms. Here, we show that acetylated and methylated histones were found to be enriched at strand switch regions of divergent gene arrays, not at convergent clusters or intra- and intergenic regions within clusters. The modified region showed a bimodular profile with two peaks centered over the 5'-regions of the gene pair flanking the strand switch region. This pattern, which demarcates polycistronic transcription units originating from bidirectional initiation sites, is likely to be common in kinetoplastid parasites as well as in other organisms with polycistronic transcription. In contrast, no acetylation was found at promoters of the highly expressed rRNA and spliced leader genes or satellite DNA or at tested retrotransposonal elements. These results reveal, for the first time, the presence of specific epigenetic marks in T. cruzi with potential implications for transcriptional regulation; they indicate that both histone modifications and bidirectional transcription are evolutionarily conserved.

Highlights

  • Tronic transcripts that are processed rapidly into individual mRNAs regulated mainly by post-transcriptional events (2)

  • Acetylated Histones H3/H4 and H3K4me3 Associate with Strand Switch Regions Separating Divergent Gene Clusters— The distribution of acetylated histones H3/H4 and H3K4me3 in 24 head-to-head (HH) and 24 tail-to-tail (TT) strand switch region (SSR) was analyzed in the T. cruzi CL Brener strain at the epimastigote stage using Chromatin Immunoprecipitation (ChIP)

  • The acetylation status of the analyzed regions did not differ among parasite strains, as similar results were obtained from ␣-acH3 ChIP experiments in other T. cruzi strains

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Summary

Introduction

Tronic transcripts that are processed rapidly into individual mRNAs regulated mainly by post-transcriptional events (2). We show a striking association of all three modifications to regions separating divergent gene clusters in the T. cruzi parasite, correlating with transcriptional initiation sites.

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