The SPARC complex defines RNAPII promoters in Trypanosoma brucei
Abstract
Kinetoplastids are a highly divergent lineage of eukaryotes with unusual mechanisms for regulating gene expression. We previously surveyed 65 putative chromatin factors in the kinetoplastid Trypanosoma brucei. Our analyses revealed that the predicted histone methyltransferase SET27 and the Chromodomain protein CRD1 are tightly concentrated at RNAPII transcription start regions (TSRs). Here we report that SET27 and CRD1, together with four previously uncharacterized constituents, form the SET27 promoter-associated regulatory complex (SPARC), which is specifically enriched at TSRs. SET27 loss leads to aberrant RNAPII recruitment to promoter sites, accumulation of polyadenylated transcripts upstream of normal transcription start sites, and conversion of some normally unidirectional promoters to bidirectional promoters. Transcriptome analysis in the absence of SET27 revealed upregulated mRNA expression in the vicinity of SPARC peaks within the main body of chromosomes in addition to derepression of genes encoding variant surface glycoproteins (VSGs) located in subtelomeric regions. These analyses uncover a novel chromatin-associated complex required to establish accurate promoter position and directionality.
Data availability
The sequencing data generated in this study can be accessed on the NCBI Gene Expression Omnibus (GEO; https://www.ncbi.nlm.nih.gov/geo/) with accession number GSE208037.The proteomics data generated in this study can be accessed on the Proteomics Identification Database (PRIDE; https://www.ebi.ac.uk/pride/) with accession number PXD036454.
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The SPARC complex defines RNAPII promoters in Trypanosoma bruceiNCBI Gene Expression Omnibus, GSE208037.
Article and author information
Author details
Funding
Medical Research Council (MR/T04702X/1)
- Desislava P Staneva
- Keith R Matthews
- Robin C Allshire
Wellcome Trust (22171)
- Keith R Matthews
Wellcome Trust (200885)
- Desislava P Staneva
- Tatsiana Auchynnikava
- Robin C Allshire
Wellcome Trust (224358)
- Desislava P Staneva
- Tatsiana Auchynnikava
- Robin C Allshire
Wellcome Trust (222516)
- Stefan Bresson
- David Tollervey
Wellcome Trust (202811)
- A Arockia Jeyaprakash
Wellcome Trust (108504)
- Juri Rappsilber
Wellcome Trust (203149)
- Christos Spanos
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Jessica K Tyler, Weill Cornell Medicine, United States
Version history
- Preprint posted: July 15, 2022 (view preprint)
- Received: September 1, 2022
- Accepted: September 27, 2022
- Accepted Manuscript published: September 28, 2022 (version 1)
- Version of Record published: October 14, 2022 (version 2)
Copyright
© 2022, Staneva et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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