Project description:Chromatin structure is a basal epigenetic mechanism that determines cellular fate by organizing the dynamic gene expression during the cell development and proliferation. The nuclear members of the evolutionarily conserved actin-related protein (ARPs) superfamily are major components of nucleosome remodelingcomplexes in the nucleus. In the human malaria parasites, Plasmodium falciparum, comparative genome analysis reveals that two canonical actins and three orthologues of ARPs including PfArp1, PfArp4, and PfArp6 are encoded in the genome of this parasite. However, little is known about the biological functions of the two nuclear PfArp4 and PfArp6 proteins. Here, by Pfarp4 gene knockdown and comparative transcriptome analysis, we uncovered that PfArp4 correlated positively with the dynamic expression of eukaryotic genes. Genome-wide distribution analysis by ChIP-seq revealed that PfArp4 protein colocalized with the histone 2A variant H2A.Z and euchromatic marker H3K9ac in the intergenic regions. Inducible downregulation of PfArp4 resulted in the depletion of H2A.Z and lower H3K9ac level at the upstream regions of eukaryotic genes, thereby repressing the transcriptional abundances. Moreover, we found a significant enrichment of PfArp4 at the flanking sites of centromeres, which likely shapes the H2A.Z-enriched centromeric chromatin microenvironment as a boundary marker. PfArp4 depletion triggered loss of H2A.Z at the entire centromere regions, and arrested the blood-stage development probably through interference with the schizogony process. Finally, PfArp6 was detected as an interactor of PfArp4 in the nucleus. Taken together, our finding indicates that the nuclear PfArp4/6regulates the cell cycles through controlling H2A.Z deposition and centromere biology, which will contribute to understanding the complex epigenetic regulation of gene expression and development of the malaria parasites.
Project description:Histone variants are key components of the epigenetic code and evolved to perform specific functions in transcriptional regulation, DNA repair, chromosome segregation and other fundamental processes. H2B.Z is a rare, apicomplexan-specific variant of histone H2B. Here we show that in Plasmodium falciparum H2B.Z localises to euchromatic intergenic regions throughout intraerythrocytic development and together with H2A.Z forms a double-variant nucleosomes subtype. These nucleosomes are enriched in promoters over 3’ intergenic regions and their occupancy generally correlates with the strength of the promoter, but not with its temporal activity. Remarkably, H2B.Z occupancy levels exhibit a clear correlation with the base-composition of the underlying DNA, raising the intriguing possibility that the extreme AT-content of the intergenic regions within the Plasmodium genome might be instructive for histone variant deposition. In summary, our data shows that the H2A.Z/H2B.Z double-variant nucleosome demarcates putative regulatory regions of the P. falciparum epigenome and likely provides a scaffold for dynamic regulation of gene expression in this deadly human pathogen. Genome-wide localization of H2B.Z has been studied at three stages of intraerythrocytic development by Illumina sequencing of chromatin-immunoprecipitated and input DNA.
Project description:Histone variants are key components of the epigenetic code and evolved to perform specific functions in transcriptional regulation, DNA repair, chromosome segregation and other fundamental processes. H2B.Z is a rare, apicomplexan-specific variant of histone H2B. Here we show that in Plasmodium falciparum H2B.Z localises to euchromatic intergenic regions throughout intraerythrocytic development and together with H2A.Z forms a double-variant nucleosomes subtype. These nucleosomes are enriched in promoters over 3’ intergenic regions and their occupancy generally correlates with the strength of the promoter, but not with its temporal activity. Remarkably, H2B.Z occupancy levels exhibit a clear correlation with the base-composition of the underlying DNA, raising the intriguing possibility that the extreme AT-content of the intergenic regions within the Plasmodium genome might be instructive for histone variant deposition. In summary, our data shows that the H2A.Z/H2B.Z double-variant nucleosome demarcates putative regulatory regions of the P. falciparum epigenome and likely provides a scaffold for dynamic regulation of gene expression in this deadly human pathogen.