The MYST family histone acetyltransferase regulates gene expression and cell cycle in malaria parasite Plasmodium falciparum

Abstract

SummaryHistone lysine acetylation, normally associated with euchromatin and active genes, is regulated by different families of histone acetyltransferases (HATs). A single Plasmodium falciparum MYST (PfMYST) HAT was expressed as a long and a short version in intraerythrocytic stages. Whereas the recombinant PfMYST expressed in prokaryotes and insect cells did not show HAT activity, recombinant PfMYST purified from the parasites exhibited a predilection to acetylate histone H4 in vitro at K5, K8, K12 and K16. Tagging PfMYST with the green fluorescent protein at the C-terminus showed that PfMYST protein was localized in both the nucleus and cytoplasm. Consistent with the importance of H4 acetylation in var gene expression, PfMYST was recruited to the active var promoter. Attempts to disrupt PfMYST were not successful, suggesting that PfMYST is essential for asexual intraerythrocytic growth. However, overexpression of the long, active or a truncated, non-active version of PfMYST by stable integration of the expression cassette in the parasite genome resulted in changes of H4 acetylation and cell cycle progression. Furthermore, parasites with PfMYST overexpression showed changes in sensitivity to DNA-damaging agents. Collectively, this study showed that PfMYST plays important roles in cellular processes such as gene activation, cell cycle control and DNA repair.

Histone H4 lysine 14 acetylation in Leishmania donovani is mediated by the MYST-family protein HAT4

“…This behaviour of HAT4, though deviant from the usual behaviour of MYST-family HATs in higher eukaryotes, which usually localize to the nucleus, is similar to what is seen in apicomplexan protozoans. MYSTfamily HATs of Toxoplasma gondii and Plasmodium falciparum have been found to localize to both the nucleus and the cytosol, and MYST-B of Toxoplasma gondii has been found to be predominantly cytosolic (Smith et al, 2005;Miao et al, 2010;Vonlaufen et al, 2010). The mammalian MYST-family HAT Tip60 has also been found to be distributed between the nucleus and the cytoplasm (Zhang et al, 2007).…”

Sex-Specific Biology of the Human Malaria Parasite Revealed from the Proteomes of Mature Male and Female Gametocytes

“…For each gene, ϳ 1 kb fragment from the C terminus end (without the stop codon) was amplified and subcloned to fuse with the GFP and 3Ј UTR region of P. berghei dhfr-ts gene, and cloned into the transfection vector pHD22Y with the human DHFR cassette (supplemental Fig. S1A, supplemental Table S1A) (64,65). Parasite transfection, drug selection and cloning were performed as described (29,65).…”

“…S1A, supplemental Table S1A) (64,65). Parasite transfection, drug selection and cloning were performed as described (29,65). Positive clones were verified by integration-specific PCR and detection of GFP expression by fluorescence microscopy (supplemental Fig.…”

Arbitrarily Accessible 3D Microfluidic Device for Combinatorial High-Throughput Drug Screening

“…To purify the Pf GCN5 HAT from the malaria parasite Plasmodium falciparum , a PTP (ProtC-TEV-ProtA) tag was added to the C-terminus of the endogenous Pf GCN5 gene in the malaria parasite as described before [36,37]. Briefly, ~1 kb fragment from the C-terminus end (without the stop codon) of Pf GCN5 was amplified and fused to the PTP tag and 3′ UTR region of the P. berghei dhfr-ts gene and cloned into the transfection vector pHD22Y with the human DHFR resistance cassette.…”

“…Briefly, ~1 kb fragment from the C-terminus end (without the stop codon) of Pf GCN5 was amplified and fused to the PTP tag and 3′ UTR region of the P. berghei dhfr-ts gene and cloned into the transfection vector pHD22Y with the human DHFR resistance cassette. Parasite transfection, drug selection, and cloning were performed as described [36,38]. Positive clones were verified by integration-specific PCR.…”