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Exploring the virulence gene interactome with CRISPR / dC as9 in the human malaria parasite
Name
msb.20209569.pdf
Description
Published version
Size
5.9 MB
Format
Adobe PDF
Checksum (MD5)
af59ed538ff1e1392f5f85bb583170a9
Author(s)
Bryant, Jessica M
Baumgarten, Sebastian
Dingli, Florent
Loew, Damarys
Sinha, Ameya
Claës, Aurélie
Preiser, Peter R
Dedon, Peter C
Scherf, Artur
Date Issued
2020
Journal
Molecular Systems Biology
Publisher
EMBO
Version
Final published version
Abstract
© 2020 The Authors. Published under the terms of the CC BY 4.0 license Mutually exclusive expression of the var multigene family is key to immune evasion and pathogenesis in Plasmodium falciparum, but few factors have been shown to play a direct role. We adapted a CRISPR-based proteomics approach to identify novel factors associated with var genes in their natural chromatin context. Catalytically inactive Cas9 (“dCas9”) was targeted to var gene regulatory elements, immunoprecipitated, and analyzed with mass spectrometry. Known and novel factors were enriched including structural proteins, DNA helicases, and chromatin remodelers. Functional characterization of PfISWI, an evolutionarily divergent putative chromatin remodeler enriched at the var gene promoter, revealed a role in transcriptional activation. Proteomics of PfISWI identified several proteins enriched at the var gene promoter such as acetyl-CoA synthetase, a putative MORC protein, and an ApiAP2 transcription factor. These findings validate the CRISPR/dCas9 proteomics method and define a new var gene-associated chromatin complex. This study establishes a tool for targeted chromatin purification of unaltered genomic loci and identifies novel chromatin-associated factors potentially involved in transcriptional control and/or chromatin organization of virulence genes in the human malaria parasite.
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
10.15252/MSB.20209569
