Plasmodium falciparum: Differential Display Analysis of Gene Expression during Gametocytogenesis

Cui, Liwang; Rzomp, Kimberly A.; Fan, Qi; Martin, Samuel K.; Williams, Jon H Sims

doi:10.1006/expr.2001.4669

Cited by 25 publications

(16 citation statements)

References 46 publications

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“…For this experiment, early trophozoites in 2 ml of complete medium were seeded in 24-well plates at 5% parasitemia and 5% hematocrit and treated with curcumin or DMSO control for 6 h at 37°C. Genomic DNA was isolated from parasite pellets using a proteinase K digestion and phenol-chloroform extraction method (8). DNA concentrations were accurately determined by dot blot analysis.…”

Section: Methodsmentioning

confidence: 99%

“…DNA concentrations were accurately determined by dot blot analysis. Blotting of serially diluted DNA onto a nylon membrane, labeling of 3D7 genomic DNA with [ 32 P]dATP by a random priming protocol, and molecular hybridization were performed as previously described (8). The hybridization intensity of each dot was measured using a phosphorimager (Molecular Dynamics, Piscataway, NJ).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Cytotoxic Effect of Curcumin on Malaria Parasite Plasmodium falciparum : Inhibition of Histone Acetylation and Generation of Reactive Oxygen Species

Cui

Miao

Cui

2007

Antimicrob Agents Chemother

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220

168

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The emergence of multidrug-resistant parasites is a major concern for malaria control, and development of novel drugs is a high priority. Curcumin, a natural polyphenolic compound, possesses diverse pharmacological properties. Among its antiprotozoan activities, curcumin was potent against both chloroquine-sensitive and -resistant Plasmodium falciparum strains. Consistent with findings in mammalian cell lines, curcumin's prooxidant activity promoted the production in P. falciparum of reactive oxygen species (ROS), whose cytotoxic effect could be antagonized by coincubation with antioxidants and ROS scavengers. Curcumin treatment also resulted in damage of both mitochondrial and nuclear DNA, probably due to the elevation of intracellular ROS. Furthermore, we have demonstrated that curcumin inhibited the histone acetyltransferase (HAT) activity of the recombinant P. falciparum general control nonderepressed 5 (PfGCN5) in vitro and reduced nuclear HAT activity of the parasite in culture. Curcumin-induced hypoacetylation of histone H3 at K9 and K14, but not H4 at K5, K8, K12, and K16, suggested that curcumin caused specific inhibition of the PfGCN5 HAT. Taken together, these results indicated that at least the generation of ROS and down-regulation of PfGCN5 HAT activity accounted for curcumin's cytotoxicity for malaria parasites.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Cytotoxic Effect of Curcumin on Malaria Parasite Plasmodium falciparum : Inhibition of Histone Acetylation and Generation of Reactive Oxygen Species

Cui

Miao

Cui

2007

Antimicrob Agents Chemother

Self Cite

220

168

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“…For most purposes, the culture was not synchronized. To isolate the parasite, the culture was treated with 0.05% saponin to lyse the red blood cell membrane and the released parasites were pelleted by centrifugation and washed twice with cold phosphate-buffered saline (15).…”

Section: Methodsmentioning

confidence: 99%

Plasmodium falciparum Histone Acetyltransferase, a Yeast GCN5 Homologue Involved in Chromatin Remodeling

2004

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The yeast transcriptional coactivator GCN5 (yGCN5), a histone acetyltransferase (HAT), is part of large multimeric complexes that are required for chromatin remodeling and transcriptional activation. Like other eukaryotes, the malaria parasite DNA is organized into nucleosomes and the genome encodes components of chromatin-remodeling complexes. Here we show that GCN5 is conserved in Plasmodium species and that the most homologous regions are within the HAT domain and the bromodomain. The Plasmodium falciparum GCN5 homologue (PfGCN5) is spliced with three introns, encoding a protein of 1,464 residues. Mapping of the ends of the PfGCN5 transcript suggests that the mRNA is 5.2 to 5.4 kb, consistent with the result from Northern analysis. Using free core histones, we determined that recombinant PfGCN5 proteins have conserved HAT activity with a substrate preference for histone H3. Using substrate-specific antibodies, we determined that both Lys-8 and -14 of H3 were acetylated by the recombinant PfGCN5. In eukaryotes, GCN5 homologues interact with yeast ADA2 homologues and form large multiprotein HAT complexes. We have identified an ADA2 homologue in P. falciparum, PfADA2. Yeast two-hybrid and in vitro binding assays verified the interactions between PfGCN5 and PfADA2, suggesting that they may be associated with each other in vivo. The conserved function of the HAT domain in PfGCN5 was further illustrated with yeast complementation experiments, which showed that the PfGCN5 region corresponding to the full-length yGCN5 could partially complement the yGCN5 deletion mutation. Furthermore, a chimera comprising the PfGCN5 HAT domain fused to the remainder of yeast GCN5 (yGCN5) fully rescued the yGCN5 deletion mutant. These data demonstrate that PfGCN5 is an authentic GCN5 family member and may exist in chromatin-remodeling complexes to regulate gene expression in P. falciparum.In eukaryotes, nuclear DNA is packaged in a hierarchical structure of 30-to 400-nm chromatin fibers composed of core histones and nonhistone proteins. Nucleosome, the primary building block of chromatin, can inhibit several processes of eukaryotic transcription, from activator binding to the promoter, to transcriptional initiation, elongation, and termination (69). The relief of nucleosome-mediated repression of transcription requires the modification of the chromatin structures, which is often accomplished by the action of two major classes of multiprotein complexes. One class, represented by the SWI/SNF complex, perturbs nucleosome structure in an ATP-dependent manner (65). The other class remodels the chromatin structure through covalent modifications (acetylation, phosphorylation, methylation, and ubiquitination) of the amino termini of the core histones in the nucleosomes (5). Within the second class, histone acetyltransferase (HAT) complexes are the best characterized (reviewed in references 8, 24, and 56). HATs acetylate lysine residues at the N-terminal tails of core histones, thereby neutralizing their positive charges. This presumably redu...

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“…Synchronization of asexual stages was performed by two rounds of sorbitol treatment as described previously (Cui et al, 2001). For the time course studies, parasites were taken at 2, 10, 18, 26, 32 h postsynchronization to represent rings, early, mid-, and late trophozoites, and schizonts as determined by microscopy.…”

Section: Parasite Culturementioning

confidence: 99%

The malaria parasite Plasmodium falciparum histones: Organization, expression, and acetylation

Miao

Fan

Cui

et al. 2006

Gene

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177

224

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Plasmodium falciparum: Differential Display Analysis of Gene Expression during Gametocytogenesis

Cited by 25 publications

References 46 publications

Cytotoxic Effect of Curcumin on Malaria Parasite Plasmodium falciparum : Inhibition of Histone Acetylation and Generation of Reactive Oxygen Species

Cytotoxic Effect of Curcumin on Malaria Parasite Plasmodium falciparum : Inhibition of Histone Acetylation and Generation of Reactive Oxygen Species

Plasmodium falciparum Histone Acetyltransferase, a Yeast GCN5 Homologue Involved in Chromatin Remodeling

The malaria parasite Plasmodium falciparum histones: Organization, expression, and acetylation

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