Quantitative chromatin proteomics reveals a dynamic histone post ...
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Received: 31 October 2016 Accepted: 8 March 2017 Published: xx xx xxxx
Quantitative chromatin proteomics reveals a dynamic histone posttranslational modification landscape that defines asexual and sexual Plasmodium falciparum parasites Nanika Coetzee1, Simone Sidoli2, Riëtte van Biljon 1, Heather Painter3, Manuel Llinás3, Benjamin A. Garcia2 & Lyn-Marie Birkholtz 1 Gene expression in Plasmodia integrates post-transcriptional regulation with epigenetic marking of active genomic regions through histone post-translational modifications (PTMs). To generate insights into the importance of histone PTMs to the entire asexual and sexual developmental cycles of the parasite, we used complementary and comparative quantitative chromatin proteomics to identify and functionally characterise histone PTMs in 8 distinct life cycle stages of P. falciparum parasites. ~500 individual histone PTMs were identified of which 106 could be stringently validated. 46 individual histone PTMs and 30 co-existing PTMs were fully quantified with high confidence. Importantly, 15 of these histone PTMs are novel for Plasmodia (e.g. H3K122ac, H3K27me3, H3K56me3). The comparative nature of the data revealed a highly dynamic histone PTM landscape during life cycle development, with a set of histone PTMs (H3K4ac, H3K9me1 and H3K36me2) displaying a unique and conserved abundance profile exclusively during gametocytogenesis (P 60% stage I; >50% stage II; >80% stage III; >90% stage IV; and >85% stage V; see Supplementary Figure S1). This allowed for the identification of all core histones typically present in P. falciparum parasites, including their variants (Fig. 1). The yields of the histone-enriched, acid-soluble nuclear protein fractions differed between the various life cycle stages (Fig. 1a) with the lowest yield observed during early asexual development (rings at 0.015 ± 0.006 ng total histones/parasite; trophozoites at 0.017 ± 0.011 ng/parasite; P > 0.8, n = 3) and a significant increase observed in schizonts (0.36 ± 0.08 ng/parasite; P
