Mapping and functional analysis of heterochromatin protein 1 phosphorylation in the malaria parasite Plasmodium falciparum

Bui, HTN; Niederwieser, I; Bird, MJ; Dai, W; Brancucci, NMB; Moes, S; Jenoe, P; Lucet, IS; Doerig, C; Voss, TS

Author(s): Bui, HTN; Niederwieser, I; Bird, MJ; Dai, W; Brancucci, NMB; Moes, S; Jenoe, P; Lucet, IS; Doerig, C; Voss, TS;
Details: Publication Year 2019-11-13,Volume 9,Issue #1,Page 16720
Journal Title: Scientific Reports
Publication Type: Journal Article
Abstract: Previous studies in model eukaryotes have demonstrated that phosphorylation of heterochromatin protein 1 (HP1) is important for dynamically regulating its various functions. However, in the malaria parasite Plasmodium falciparum both the function of HP1 phosphorylation and the identity of the protein kinases targeting HP1 are still elusive. In order to functionally analyze phosphorylation of P. falciparum HP1 (PfHP1), we first mapped PfHP1 phosphorylation sites by liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis of native PfHP1, which identified motifs from which potential kinases could be predicted; in particular, several phosphorylated residues were embedded in motifs rich in acidic residues, reminiscent of targets for P. falciparum casein kinase 2 (PfCK2). Secondly, we tested recombinant PfCK2 and a number of additional protein kinases for their ability to phosphorylate PfHP1 in in vitro kinase assays. These experiments validated our prediction that PfHP1 acts as a substrate for PfCK2. Furthermore, LC-MS/MS analysis showed that PfCK2 phosphorylates three clustered serine residues in an acidic motif within the central hinge region of PfHP1. To study the role of PfHP1 phosphorylation in live parasites we used CRISPR/Cas9-mediated genome editing to generate a number of conditional PfHP1 phosphomutants based on the DiCre/LoxP system. Our studies revealed that neither PfCK2-dependent phosphorylation of PfHP1, nor phosphorylation of the hinge domain in general, affect PfHP1's ability to localize to heterochromatin, and that PfHP1 phosphorylation in this region is dispensable for the proliferation of P. falciparum blood stage parasites.
Publisher: NPG
Research Division(s): Chemical Biology
PubMed ID: 31723180
Publisher's Version: https://doi.org/10.1038/s41598-019-53325-9
Open Access at Publisher's Site: https://doi.org/10.1038/s41598-019-53325-9
Terms of Use/Rights Notice: Refer to copyright notice on published article.
Documents: Bui HTN et al_Scientific Reports_2019.pdf - (3.28MB, application/pdf)

Creation Date: 2019-11-20 02:45:16

Last Modified: 2019-11-21 09:43:05