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PXD061709

PXD061709 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitlePrecision-edited histone tails perturb polycistronic gene expression controls in trypanosomes
DescriptionTranscription of protein coding genes in trypanosomatids is atypical and almost exclusively polycistronic. In Trypanosoma brucei, approximately 150 polycistrons, and 8000 genes, are constitutively transcribed by RNA polymerase II, while a smaller subset can be transcribed by RNA polymerase I. RNA polymerase II promoters are unconventional and characterised by regions of chromatin enriched for histones with specific patterns of post-translational modification on their highly divergent N-terminal tails. To investigate the roles of histone tail-residues in gene expression control in T. brucei, we engineered strains exclusively expressing novel mutant histones. We used an inducible CRISPR-Cas9 system to delete >40 native copies of histone H4, complementing the tandem arrays with a single ectopic H4 gene. The resulting ‘histoneH4’ strains were validated using whole-genome sequencing and transcriptome analysis. We then performed saturation mutagenesis of six histone H4 N-terminal tail lysine (K) residues and used multiplex amplicon-seq to profile the relative fitness of 384 distinct precision edited mutants. H4K10 mutations were not tolerated, but we could derive a panel of nineteen strains exclusively expressing novel H4K4 or H4K14 mutants. Both proteomic and transcriptomic analysis of H4K4Q mutants revealed significantly reduced expression of genes adjacent to RNA polymerase II promoters, where the glutamine (Q) mutation mimics an abnormally high level of acetylation. Thus, we present direct evidence for polycistronic expression control by histone H4 N-terminal tails in trypanosomes.
HostingRepositoryPRIDE
AnnounceDate2025-03-11
AnnouncementXMLSubmission_2025-03-11_11:36:02.977.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterMichele Tinti
SpeciesList scientific name: Trypanosoma brucei; NCBI TaxID: 5691;
ModificationListiodoacetamide derivatized residue
InstrumentLTQ Orbitrap
Dataset History
RevisionDatetimeStatusChangeLog Entry
02025-03-11 09:11:48ID requested
12025-03-11 11:36:03announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: trypanosomes H4 T. brucei polycistronic expression control
Contact List
David Horn
contact affiliationWellcome Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, UK
contact emaild.horn@dundee.ac.uk
lab head
Michele Tinti
contact affiliationDundee University
contact emailm.tinti@dundee.ac.uk
dataset submitter
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